SUBSTITUTED 1,2,3,4-TETRAHYDROCYCLOPENTA[b]INDOL-3-YL)ACETIC ACID DERIVATIVES USEFUL IN THE TREATMENT OF AUTOIMMUNE AND INFLAMMATORY DISORDERS

ABSTRACT

The present invention relates to certain substituted 1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetic acid derivatives of Formula (Ia) and pharmaceutically acceptable salts thereof, which exhibit useful pharmacological properties, for example, as agonists of the S1P1 receptor. Also provided by the present invention are pharmaceutical compositions containing compounds of the invention, and methods of using the compounds and compositions of the invention in the treatment of S1P1 receptor-associated disorders, for example, psoriasis, rheumatoid arthritis, Crohn&#39;s disease, transplant rejection, multiple sclerosis, systemic lupus erythematosus, ulcerative colitis, type I diabetes, acne, microbial infections or diseases and viral infections or diseases.

FIELD OF THE INVENTION

The present invention relates to certain substituted1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetic acid derivatives ofFormula (Ia) and pharmaceutically acceptable salts thereof, whichexhibit useful pharmacological properties, for example, as agonists ofthe S1P1 receptor. Also provided by the present invention arepharmaceutical compositions containing compounds of the invention, andmethods of using the compounds and compositions of the invention in thetreatment of S1P1 associated disorders, for example, psoriasis,rheumatoid arthritis, Crohn's disease, transplant rejection, multiplesclerosis, systemic lupus erythematosus, ulcerative colitis, type Idiabetes, acne, microbial infections or diseases and viral infections ordiseases.

BACKGROUND OF THE INVENTION

The present invention relates to compounds that are S1P1 receptoragonists having at least immunosuppressive, anti-inflammatory and/orhemostatic activities, e.g. by virtue of modulating leukocytetrafficking, sequestering lymphocytes in secondary lymphoid tissues,and/or enhancing vascular integrity.

The present application is in part focused on addressing an unmet needfor immunosuppressive agents such as may be orally available which havetherapeutic efficacy for at least autoimmune diseases and disorders,inflammatory diseases and disorders (e.g., acute and chronicinflammatory conditions), transplant rejection, cancer, and/orconditions that have an underlying defect in vascular integrity or thatare associated with angiogenesis such as may be pathologic (e.g., as mayoccur in inflammation, tumor development and atherosclerosis) with fewerside effects such as the impairment of immune responses to systemicinfection.

The sphingosine-1-phosphate (S1P) receptors 1-5 constitute a family of Gprotein-coupled receptors with a seven-transmembrane domain. Thesereceptors, referred to as S1P1 to S1P5 (formerly termed endothelialdifferentiation gene (EDG) receptor-1, -5, -3, -6 and -8, respectively;Chun et al., Pharmacological Reviews, 54:265-269, 2002), are activatedvia binding by sphingosine-1-phosphate, which is produced by thesphingosine kinase-catalyzed phosphorylation of sphingosine. S1P1, S1P4and S1P5 receptors activate Gi but not Gq, whereas S1P2 and S1P3receptors activate both Gi and Gq. The S1P3 receptor, but not the S1P1receptor, responds to an agonist with an increase in intracellularcalcium.

S1P receptor agonists having agonist activity on the S1P1 receptor havebeen shown to rapidly and reversibly induce lymphopenia (also referredto as peripheral lymphocyte lowering (PLL); Hale et al., Bioorg. Med.Chem. Lett., 14:3351-3355, 2004). This is attended by clinically usefulimmunosuppression by virtue of sequestering T- and B-cells in secondarylymphoid tissue (lymph nodes and Peyer's patches) and thus apart fromsites of inflammation and organ grafts (Rosen et al., Immunol. Rev.,195:160-177, 2003; Schwab et al., Nature Immunol., 8:1295-1301, 2007).This lymphocyte sequestration, for example in lymph nodes, is thought tobe a consequence of concurrent agonist-driven functional antagonism ofthe S1P1 receptor on T-cells (whereby the ability of S1P to mobilizeT-cell egress from lymph nodes is reduced) and persistent agonism of theS1P1 receptor on lymph node endothelium (such that barrier functionopposing transmigration of lymphocytes is increased) (Matloubian et al.,Nature, 427:355-360, 2004; Baumruker et al., Expert Opin. Investig.Drugs, 16:283-289, 2007). It has been reported that agonism of the S1P1receptor alone is sufficient to achieve lymphocyte sequestration (Sannaet al., J Biol Chem., 279:13839-13848, 2004) and that this occurswithout impairment of immune responses to systemic infection (Brinkmannet al., Transplantation, 72:764-769, 2001; Brinkmann et al., TransplantProc., 33:530-531, 2001).

That agonism of endothelial S1P1 receptors has a broader role inpromoting vascular integrity is supported by work implicating the S1P1receptor in capillary integrity in mouse skin and lung (Sanna et al.,Nat Chem Biol., 2:434-441, 2006). Vascular integrity can be compromisedby inflammatory processes, for example as may derive from sepsis, majortrauma and surgery so as to lead to acute lung injury or respiratorydistress syndrome (Johan Groeneveld, Vascul. Pharmacol., 39:247-256,2003).

An exemplary S1P receptor agonist having agonist activity on the S1P1receptor is FTY720 (fingolimod), an immunosuppressive agent currently inclinical trials (Martini et al., Expert Opin. Investig. Drugs,16:505-518, 2007). FTY720 acts as a prodrug which is phosphorylated invivo; the phosphorylated derivative is an agonist for S1P1, S1P3, S1P4and S1P5 receptors (but not the S1P2 receptor) (Chiba, Pharmacology &Therapeutics, 108:308-319, 2005). FTY720 has been shown to rapidly andreversibly induce lymphopenia (also referred to as peripheral lymphocytelowering (PLL); Hale et al., Bioorg. Med. Chem. Lett., 14:3351-3355,2004). This is attended by clinically useful immunosuppression by virtueof sequestering T- and B-cells in secondary lymphoid tissue (lymph nodesand Peyer's patches) and thus apart from sites of inflammation and organgrafts (Rosen et al., Immunol. Rev., 195:160-177, 2003; Schwab et al.,Nature Immunol., 8:1295-1301, 2007).

In clinical trials, FTY720 elicited an adverse event (i.e., transientasymptomatic bradycardia) due to its agonism of the S1P3 receptor (Buddeet al., J. Am. Soc. Nephrol., 13:1073-1083, 2002; Sanna et al., J. Biol.Chem., 279:13839-13848, 2004; Ogawa et al., BBRC, 361:621-628, 2007).

FTY720 has been reported to have therapeutic efficacy in at least: a ratmodel for autoimmune myocarditis and a mouse model for acute viralmyocarditis (Kiyabayashi et al., J. Cardiovasc. Pharmacol., 35:410-416,2000; Miyamoto et al., J. Am. Coll. Cardiol., 37:1713-1718, 2001); mousemodels for inflammatory bowel disease including colitis (Mizushima etal., Inflamm. Bowel Dis., 10:182-192, 2004; Deguchi et al., OncologyReports, 16:699-703, 2006; Fujii et al., Am. J. Physiol. Gastrointest.Liver Physiol., 291:G267-G274, 2006; Daniel et al., J. Immunol.,178:2458-2468, 2007); a rat model for progressive mesangioproliferativeglomerulonephritis (Martini et al., Am. J. Physiol. Renal Physiol.,292:F1761-F1770, 2007); a mouse model for asthma, suggested to beprimarily through the S1P1 receptor on the basis of work using the S1P1receptor agonist SEW2871 (Idzko et al, J. Clin. Invest., 116:2935-2944,2006); a mouse model for airway inflammation and induction of bronchialhyperresponsiveness (Sawicka et al., J. Immunol., 171; 6206-6214, 2003);a mouse model for atopic dermatitis (Kohno et al., Biol. Pharm. Bull.,27:1392-1396, 2004); a mouse model for ischemia-reperfusion injury(Kaudel et al., Transplant. Proc, 39:499-502, 2007); a mouse model forsystemic lupus erythematosus (SLE) (Okazaki et al., J. Rheumatol.,29:707-716, 2002; Herzinger et al, Am. J. Clin. Dermatol., 8:329-336,2007); rat models for rheumatoid arthritis (Matsuura et al., Int. J.Imnzunopharmacol., 22:323-331, 2000; Matsuura et al., Inflamm. Res.,49:404-410, 2000); a rat model for autoimmune uveitis (Kurose et al.,Exp. Eye Res., 70:7-15, 2000); mouse models for type I diabetes (Fu etal, Transplantation, 73:1425-1430, 2002; Maki et al., Transplantation,74:1684-1686, 2002; Yang et al., Clinical Immunology, 107:30-35, 2003;Maki et al., Transplantation, 79:1051-1055, 2005); mouse models foratherosclerosis (Nofer et al., Circulation, 115:501-508, 2007; Keul etal., Arterioscler. Thromb. Vase. Biol., 27:607-613, 2007); a rat modelfor brain inflammatory reaction following traumatic brain injury (TBI)(Zhang et al., J. Cell. Mol. Med., 11:307-314, 2007); and mouse modelsfor graft coronary artery disease and graft-versus-host disease (GVHD)(Hwang et al., Circulation, 100:1322-1329, 1999; Taylor et al., Blood,110:3480-3488, 2007). In vitro results suggest that FTY720 may havetherapeutic efficacy for β-amyloid-related inflammatory diseasesincluding Alzheimer's disease (Kaneider et al., FASEB J., 18:309-311,2004). KRP-203, an S1P receptor agonist having agonist activity on theS1P1 receptor, has been reported to have therapeutic efficacy in a ratmodel for autoimmune myocarditis (Ogawa et al., BBRC, 361:621-628,2007). Using the S1P1 receptor agonist SEW2871, it has been shown thatagonism of endothelial S1P1 receptors prevents proinflammatorymonocyte/endothelial interactions in type I diabetic vascularendothelium (Whetzel et al., Circ. Res., 99:731-739, 2006) and protectsthe vasculature against TNFα-mediated monocyte/endothelial interactions(Bolick et al., Arterioscler. Thromb. Vasc. Biol., 25:976-981, 2005).

Additionally, FTY720 has been reported to have therapeutic efficacy inexperimental autoimmune encephalomyelitis (EAE) in rats and mice, amodel for human multiple sclerosis (Brinkmann et al., J. Biol. Chem.,277:21453-21457, 2002; Fujino et al., J. Pharmacol. Exp. Ther.,305:70-77, 2003; Webb et al., J. Neuroimmunol., 153:108-121, 2004;Rausch et al., J. Magn. Reson. Imaging, 20:16-24, 2004; Kataoka et al.,Cellular & Molecular Immunology, 2:439-448, 2005; Brinkmann et al.,Pharmacology & Therapeutics, 115:84-105, 2007; Baumruker et al., ExpertOpin. Investig. Drugs, 16:283-289, 2007; Balatoni et al., Brain ResearchBulletin, 74:307-316, 2007). Furthermore, FTY720 has been found to havetherapeutic efficacy for multiple sclerosis in clinical trials. In PhaseII clinical trials for relapsing-remitting multiple sclerosis, FTY720was found to reduce the number of lesions detected by magnetic resonanceimaging (MRI) and clinical disease activity in patients with multiplesclerosis (Kappos et al., N. Engl. J. Med., 355:1124-1140, 2006; Martiniet al., Expert Opin. Investig. Drugs, 16:505-518, 2007; Zhang et al.,Mini-Reviews in Medicinal Chemistry, 7:845-850, 2007; Brinkmann,Pharmacology & Therapeutics, 115:84-105, 2007). FTY720 is currently inPhase III studies of remitting-relapsing multiple sclerosis (Brinkmann,Pharmacology & Therapeutics, 115:84-105, 2007; Baumruker et al., Expert.Opin. Investig. Drugs, 16:283-289, 2007; Dev et al., Pharmacology andTherapeutics, 117:77-93, 2008).

Recently, FTY720 has been reported to have anti-viral activity. Specificdata has been presented in the lymphocytic choriomeningitis virus (LCMV)mouse model, wherein the mice were infected with either the Armstrong orthe clone 13 strain of LCMV (Premenko-Lanier et al., Nature, 454, 894,2008).

FTY720 has been reported to impair migration of dendritic cells infectedwith Francisella tularensis to the mediastinal lymph node, therebyreducing the bacterial colonization of it. Francisella tularensis isassociated with tularemia, ulceroglandular infection, respiratoryinfection and a typhoidal disease (E. Bar-Haim et al, PLoS Pathogens,4(11): e1000211. doi:10.1371/journal.ppat.1000211, 2008).

It has also been recently reported that a short-term high dose of FTY720rapidly reduced ocular infiltrates in experimental autoimmuneuveoretinitis. When given in the early stages of ocular inflammation,FTY720 rapidly prevented retinal damage. It was reported to not onlyprevent infiltration of target organs, but also reduce existinginfiltration (Raveney et al., Arch. Ophthalmol. 126(10), 1390, 2008).

It has been reported that treatment with FTY720 relievedovariectomy-induced osteoporosis in mice by reducing the number ofmature osteoclasts attached to the bone surface. The data providedevidence that S1P controlled the migratory behaviour of osteoclastprecursors, dynamically regulating bone mineral homeostasis (Ishii etal., Nature, advance online publication, 8 Feb. 2009,doi:10.1038/nature07713).

Agonism of the S1P1 receptor has been implicated in enhancement ofsurvival of oligodendrocyte progenitor cells. Survival ofoligodendrocyte progenitor cells is a required component of theremyelination process. Remyelination of multiple sclerosis lesions isconsidered to promote recovery from clinical relapses. (Miron et al.,Ann. Neurol., 63:61-71, 2008; Coelho et al., J. Pharmacol. Exp. Ther.,323:626-635, 2007; Dev et al., Pharmacology and Therapeutics, 117:77-93,2008). It also has been shown that the S1P1 receptor plays a role inplatelet-derived growth factor (PDGF)-induced oligodendrocyte progenitorcell mitogenesis (Jung et al., Glia, 55:1656-1667, 2007).

Agonism of the S1P1 receptor has also been reported to mediate migrationof neural stem cells toward injured areas of the central nervous system(CNS), including in a rat model of spinal cord injury (Kimura et al.,Stem Cells, 25:115-124, 2007).

Agonism of the S1P1 receptor has been implicated in the inhibition ofkeratinocyte proliferation (Sauer et al., J. Biol. Chem.,279:38471-38479, 2004), consistent with reports that S1P inhibitskeratinocyte proliferation (Kim et al., Cell Signal, 16:89-95, 2004).The hyperproliferation of keratinocytes at the entrance to the hairfollicle, which can then become blocked, and an associated inflammationare significant pathogenetic factors of acne (Koreck et al.,Dermatology, 206:96-105, 2003; Webster, Cutis, 76:4-7, 2005).

FTY720 has been reported to have therapeutic efficacy in inhibitingpathologic angiogenesis, such as that as may occur in tumor development.Inhibition of angiogenesis by FTY720 is thought to involve agonism ofthe S1P1 receptor (Oo et al., J. Biol. Chem., 282; 9082-9089, 2007;Schmid et al., J. Cell Biochem., 101:259-270, 2007). FTY720 has beenreported to have therapeutic efficacy for inhibiting primary andmetastatic tumor growth in a mouse model of melanoma (LaMontagne et al.,Cancer Res., 66:221-231, 2006). FTY720 has been reported to havetherapeutic efficacy in a mouse model for metastatic hepatocellularcarcinoma (Lee et al., Clin. Cancer Res., 11:84588466, 2005).

It has been reported that oral administration of FTY720 to mice potentlyblocked VEGF-induced vascular permeability, an important processassociated with angiogenesis, inflammation, and pathological conditionssuch as sepsis, hypoxia, and solid tumor growth (T Sanchez et al, J.Biol. Chem., 278(47), 47281-47290, 2003).

Cyclosporin A and FK506 (calcineurin inhibitors) are drugs used toprevent rejection of transplanted organs. Although they are effective indelaying or suppressing transplant rejection, classicalimmunosuppressants such as cyclosporin A and FK506 are known to causeseveral undesirable side effects including nephrotoxicity,neurotoxicity, β-cell toxicity and gastrointestinal discomfort. There isan unmet need in organ transplantation for an immunosuppressant withoutthese side effects which is effective as a monotherapy or in combinationwith a classical immunosuppressant for inhibiting migration of, e.g.,alloantigen-reactive T-cells to the grafted tissue, thereby prolonginggraft survival.

FTY720 has been shown to have therapeutic efficacy in transplantrejection both as a monotherapy and in synergistic combination with aclassical immunosuppressant, including cyclosporin A, FK506 and RAD (anmTOR inhibitor). It has been shown that, unlike the classicalimmunosuppressants cyclosporin A, FK506 and RAD, FTY720 has efficacy forprolonging graft survival without inducing general immunosuppression,and this difference in drug action is believed to be relevant to thesynergism observed for the combination (Brinkmann et al., TransplantProc., 33:530-531, 2001; Brinkmann et al., Transplantation, 72:764-769,2001).

Agonism of the S1P1 receptor has been reported to have therapeuticefficacy for prolonging allograft survival in mouse and rat skinallograft models (Lima et al., Transplant Proc., 36:1015-1017, 2004; Yanet al., Bioorg. & Med. Chem. Lett., 16:3679-3683, 2006). FTY720 has beenreported to have therapeutic efficacy for prolonging allograft survivalin a rat cardiac allograft model (Suzuki et al., Transpl. Immunol.,4:252-255, 1996). FTY720 has been reported to act synergistically withcyclosporin A to prolong rat skin allograft survival (Yanagawa et al.,J. Immunol., 160:5493-5499, 1998), to act synergistically withcyclosporin A and with FK506 to prolong rat cardiac allograft survival,and to act synergistically with cyclosporin A to prolong canine renalallograft survival and monkey renal allograft survival (Chiba et al.,Cell Mol. Biol., 3:11-19, 2006). KRP-203, an S1P receptor agonist hasbeen reported to have therapeutic efficacy for prolonging allograftsurvival in a rat skin allograft model and both as monotherapy and insynergistic combination with cyclosporin A in a rat cardiac allograftmodel (Shimizu et al., Circulation, 111:222-229, 2005). KRP-203 also hasbeen reported to have therapeutic efficacy in combination withmycophenolate mofetil (MMF; a prodrug for which the active metabolite ismycophenolic acid, an inhibitor of purine biosynthesis) for prolongingallograft survival both in a rat renal allograft model and in a ratcardiac allograft model (Suzuki et al., J. Heart Lung Transplant,25:302-209, 2006; Fujishiro et al., J. Heart Lung Transplant,25:825-833, 2006). It has been reported that an agonist of the S1P1receptor, AUY954, in combination with a subtherapeutic dose of RAD001(Certican/Everolimus, an mTOR inhibitor) can prolong rat cardiacallograft survival (Pan et al., Chemistry & Biology, 13:1227-1234,2006). In a rat small bowel allograft model, FTY720 has been reported toact synergistically with cyclosporin A to prolong small bowel allograftsurvival (Sakagawa et al., Transpl. Immunol., 13:161-168, 2004). FTY720has been reported to have therapeutic efficacy in a mouse islet graftmodel (Fu et al., Transplantation, 73:1425-1430, 2002; Liu et al.,Microsurgery, 27:300-304; 2007) and in a study using human islet cellsto evidence no detrimental effects on human islet function (Truong etal., American Journal of Transplantation, 7:2031-2038, 2007).

FTY720 has been reported to reduce the nociceptive behavior in thespared nerve injury model for neuropathic pain which does not depend onprostaglandin synthesis (O. Costo et al, Journal of Cellular andMolecular Medicine 12(3), 995-1004, 2008).

FTY720 has been reported to impair initiation of murine contacthypersensitivity (CHS). Adoptive transfer of immunized lymph node cellsfrom mice treated with FTY720 during the sensitization phase wasvirtually incapable of inducing CHS response in recipients (D. Nakashimaet al., J. Investigative Dermatology (128(12), 2833-2841, 2008).

It has been reported that prophylactic oral administration of FTY720 (1mg/kg, three times a week), completely prevented the development ofexperimental autoimmune myasthenia gravis (EAMG) in C57BL/6 mice (T.Kohono et al, Biological & Pharmaceutical Bulletin, 28(4), 736-739,2005).

In one embodiment, the present invention encompasses compounds which areagonists of the S1P1 receptor having selectivity over the S1P3 receptor.The S1P3 receptor, and not the S1P1 receptor, has been directlyimplicated in bradycardia (Sanna et al., J. Biol. Chem.,279:13839-13848, 2004). An S1P1 receptor agonist selective over at leastthe S1P3 receptor has advantages over current therapies by virtue of anenhanced therapeutic window, allowing better tolerability with higherdosing and thus improving efficacy as therapy. The present inventionencompasses compounds which are agonists of the S1P1 receptor and whichexhibit no or substantially no activity for bradycardia.

S1P1 receptor agonists are useful to treat or prevent conditions wheresuppression of the immune system or agonism of the S1P1 receptor is inorder, such as diseases and disorders mediated by lymphocytes,transplant rejection, autoimmune diseases and disorders, inflammatorydiseases and disorders, and conditions that have an underlying defect invascular integrity or that relate to angiogenesis such as may bepathologic.

In one embodiment, the present invention encompasses compounds which areagonists of the S1P1 receptor having good overall physical propertiesand biological activities and having an effectiveness that issubstantially at least that of prior compounds with activity at the S1P1receptor.

Citation of any reference throughout this application is not to beconstrued as an admission that such reference is prior art to thepresent application.

SUMMARY OF THE INVENTION

The present invention encompasses compounds of Formula (Ia) andpharmaceutically acceptable salts, solvates and hydrates thereof:

wherein:

-   -   m is 1 or 2;    -   n is 1 or 2;    -   Y is N or CR¹;    -   Z is N or CR⁴; and    -   R¹, R², R³ and R⁴ are each independently selected from the group        consisting of H, C₁-C₆ alkoxy, C₁-C₆ alkyl, C₁-C₆ alkylamino,        C₁-C₆ alkylsulfonyl, C₁-C₆ alkylthio, carboxamide, cyano, C₃-C₇        cycloalkoxy, C₃-C₇ cycloalkyl, C₁-C₆ haloalkoxy, C₁-C₆        haloalkyl, halogen, heteroaryl and heterocyclyl, wherein said        C₁-C₆ alkyl and C₁-C₆ alkoxy are each optionally substituted        with one or two substituents selected from C₃-C₇ cycloalkyl and        halogen.

In some embodiments, the present invention encompasses compounds ofFormula (Ia) and pharmaceutically acceptable salts, solvates andhydrates thereof:

wherein:

-   -   m is 1 or 2;    -   n is 1 or 2;    -   Y is N or CR¹;    -   Z is N or CR⁴; and    -   R¹, R², R³ and R⁴ are each independently selected from the group        consisting of H, C₁-C₆ alkoxy, C₁-C₆ alkyl, C₁-C₆ alkylamino,        C₁-C₆ alkylsulfonyl, C₁-C₆ alkylthio, carboxamide, cyano, C₃-C₇        cycloalkoxy, C₃-C₇ cycloalkyl, C₁-C₆ haloalkoxy, C₁-C₆        haloalkyl, halogen, heteroaryl and heterocyclyl, wherein said        C₁-C₆ alkyl and C₁-C₆ alkoxy are each optionally substituted        with one C₃-C₇ cycloalkyl group.

The present invention encompasses compounds which are S1P1 receptoragonists having at least immunosuppressive, anti-inflammatory and/orhemostatic activities, e.g. by virtue of modulating leukocytetrafficking, sequestering lymphocytes in secondary lymphoid tissues,and/or enhancing vascular integrity.

S1P1 receptor agonists are useful to treat or prevent conditions wheresuppression of the immune system or agonism of the S1P1 receptor is inorder, such as diseases and disorders mediated by lymphocytes,transplant rejection, autoimmune diseases and disorders, inflammatorydiseases and disorders (e.g., acute and chronic inflammatoryconditions), cancer, and conditions that have an underlying defect invascular integrity or that are associated with angiogenesis such as maybe pathologic (e.g., as may occur in inflammation, tumor development andatherosclerosis). Such conditions where suppression of the immune systemor agonism of the S1P1 receptor is in order include diseases anddisorders mediated by lymphocytes, conditions that have an underlyingdefect in vascular integrity, autoimmune diseases and disorders,inflammatory diseases and disorders (e.g., acute and chronicinflammatory conditions), acute or chronic rejection of cells, tissue orsolid organ grafts, arthritis including psoriatic arthritis andrheumatoid arthritis, diabetes including type I diabetes, demyelinatingdisease including multiple sclerosis, ischemia-reperfusion injuryincluding renal and cardiac ischemia-reperfusion injury, inflammatoryskin disease including psoriasis, atopic dermatitis and acne,hyperproliferative skin disease including acne, inflammatory boweldisease including Crohn's disease and ulcerative colitis, systemic lupuserythematosis, asthma, uveitis, myocarditis, allergy, atherosclerosis,brain inflammation including Alzheimer's disease and brain inflammatoryreaction following traumatic brain injury, central nervous systemdisease including spinal cord injury or cerebral infarction, pathologicangiogenesis including as may occur in primary and metastatic tumorgrowth, rheumatoid arthritis, diabetic retinopathy and atherosclerosis,cancer, chronic pulmonary disease, acute lung injury, acute respiratorydisease syndrome, sepsis and the like.

One aspect of the present invention pertains to pharmaceuticalcompositions comprising a compound of the present invention and apharmaceutically acceptable carrier.

One aspect of the present invention pertains to pharmaceuticalcompositions comprising a compound of the present invention, a salt, ahydrate or solvate or a crystalline form and a pharmaceuticallyacceptable carrier.

One aspect of the present invention pertains to methods for treating adisorder associated with the S1P1 receptor in an individual comprisingadministering to the individual in need thereof a therapeuticallyeffective amount of a compound of the present invention or apharmaceutical composition thereof.

One aspect of the present invention pertains to methods for treating anS1P1 receptor-associated disorder in an individual comprisingadministering to the individual in need thereof a therapeuticallyeffective amount of a compound of the present invention or apharmaceutical composition thereof.

One aspect of the present invention pertains to methods for treating anS1P1 receptor-associated disorder associated with the S1P1 receptor inan individual comprising administering to the individual in need thereofa therapeutically effective amount of a compound of the presentinvention, a salt, a hydrate or solvate, a crystalline form, or apharmaceutical composition thereof.

One aspect of the present invention pertains to methods for treating adisorder associated with the S1P1 receptor in an individual comprisingadministering to the individual in need thereof a therapeuticallyeffective amount of a compound of the present invention or apharmaceutical composition thereof, wherein said disorder associatedwith the S1P1 receptor is selected from the group consisting of: adisease or disorder mediated by lymphocytes, an autoimmune disease ordisorder, an inflammatory disease or disorder, cancer, psoriasis,rheumatoid arthritis, Crohn's disease, transplant rejection, multiplesclerosis, systemic lupus erythematosus, ulcerative colitis, type Idiabetes and acne.

One aspect of the present invention pertains to methods for treating adisorder associated with the S1P1 receptor in an individual comprisingadministering to the individual in need thereof a therapeuticallyeffective amount of a compound of the present invention, a salt, ahydrate or solvate, a crystalline form, or a pharmaceutical compositionthereof, wherein said disorder associated with the S1P1 receptor isselected from the group consisting of: a disease or disorder mediated bylymphocytes, an autoimmune disease or disorder, an inflammatory diseaseor disorder, cancer, psoriasis, rheumatoid arthritis, Crohn's disease,transplant rejection, multiple sclerosis, systemic lupus erythematosus,ulcerative colitis, type I diabetes and acne.

One aspect of the present invention pertains to methods for treating adisease or disorder mediated by lymphocytes in an individual comprisingadministering to the individual in need thereof a therapeuticallyeffective amount of a compound of the present invention or apharmaceutical composition thereof.

One aspect of the present invention pertains to methods for treating anautoimmune disease or disorder in an individual comprising administeringto the individual in need thereof a therapeutically effective amount ofa compound of the present invention or a pharmaceutical compositionthereof.

One aspect of the present invention pertains to methods for treating aninflammatory disease or disorder in an individual comprisingadministering to the individual in need thereof a therapeuticallyeffective amount of a compound of the present invention or apharmaceutical composition thereof.

One aspect of the present invention pertains to methods for treatingcancer in an individual comprising administering to the individual inneed thereof a therapeutically effective amount of a compound of thepresent invention or a pharmaceutical composition thereof.

One aspect of the present invention pertains to methods for treating adisorder in an individual comprising administering to the individual inneed thereof a therapeutically effective amount of a compound of thepresent invention or a pharmaceutical composition thereof, wherein saiddisorder is selected from the group consisting of psoriasis, rheumatoidarthritis, Crohn's disease, transplant rejection, multiple sclerosis,systemic lupus erythematosus, ulcerative colitis, type I diabetes andacne.

One aspect of the present invention pertains to methods for treatingpsoriasis in an individual comprising administering to the individual inneed thereof a therapeutically effective amount of a compound of thepresent invention or a pharmaceutical composition thereof.

One aspect of the present invention pertains to methods for treatingrheumatoid arthritis in an individual comprising administering to theindividual in need thereof a therapeutically effective amount of acompound of the present invention or a pharmaceutical compositionthereof.

One aspect of the present invention pertains to methods for treatingCrohn's disease in an individual comprising administering to theindividual in need thereof a therapeutically effective amount of acompound of the present invention or a pharmaceutical compositionthereof.

One aspect of the present invention pertains to methods for treatingtransplant rejection in an individual comprising administering to theindividual in need thereof a therapeutically effective amount of acompound of the present invention or a pharmaceutical compositionthereof.

One aspect of the present invention pertains to methods for treatingmultiple sclerosis in an individual comprising administering to theindividual in need thereof a therapeutically effective amount of acompound of the present invention or a pharmaceutical compositionthereof.

One aspect of the present invention pertains to methods for treatingsystemic lupus erythematosus in an individual comprising administeringto the individual in need thereof a therapeutically effective amount ofa compound of the present invention or a pharmaceutical compositionthereof.

One aspect of the present invention pertains to methods for treatingulcerative colitis in an individual comprising administering to theindividual in need thereof a therapeutically effective amount of acompound of the present invention or a pharmaceutical compositionthereof.

One aspect of the present invention pertains to methods for treatingtype I diabetes in an individual comprising administering to theindividual in need thereof a therapeutically effective amount of acompound of the present invention or a pharmaceutical compositionthereof.

One aspect of the present invention pertains to methods for treatingacne in an individual comprising administering to the individual in needthereof a therapeutically effective amount of a compound of the presentinvention or a pharmaceutical composition thereof.

One aspect of the present invention pertains to methods for treating adisorder associated with the S1P1 receptor in an individual comprisingadministering to the individual in need thereof a therapeuticallyeffective amount of a compound of the present invention or apharmaceutical composition thereof, wherein said disorder associatedwith the S1P1 receptor is a microbial infection or disease or a viralinfection or disease.

One aspect of the present invention pertains to methods for treating adisorder associated with the S1P1 receptor in an individual comprisingadministering to the individual in need thereof a therapeuticallyeffective amount of a compound of the present invention, a salt, ahydrate or solvate, a crystalline form, or a pharmaceutical compositionthereof, wherein said disorder associated with the S1P1 receptor is amicrobial infection or disease or a viral infection or disease.

One aspect of the present invention pertains to methods for treatinggastritis in an individual comprising administering to the individual inneed thereof a therapeutically effective amount of a compound of thepresent invention or a pharmaceutical composition thereof.

One aspect of the present invention pertains to methods for treatingpolymyositis in an individual comprising administering to the individualin need thereof a therapeutically effective amount of a compound of thepresent invention or a pharmaceutical composition thereof.

One aspect of the present invention pertains to methods for treatingthyroiditis in an individual comprising administering to the individualin need thereof a therapeutically effective amount of a compound of thepresent invention or a pharmaceutical composition thereof.

One aspect of the present invention pertains to methods for treatingvitiligo in an individual comprising administering to the individual inneed thereof a therapeutically effective amount of a compound of thepresent invention or a pharmaceutical composition thereof.

One aspect of the present invention pertains to methods for treatinghepatitis in an individual comprising administering to the individual inneed thereof a therapeutically effective amount of a compound of thepresent invention or a pharmaceutical composition thereof.

One aspect of the present invention pertains to methods for treatingbiliary cirrhosis in an individual comprising administering to theindividual in need thereof a therapeutically effective amount of acompound of the present invention or a pharmaceutical compositionthereof.

One aspect of the present invention pertains to the use of compounds ofthe present invention in the manufacture of a medicament for thetreatment of an S1P1 receptor-associated disorder.

One aspect of the present invention pertains to the use of compounds ofthe present invention, a salt, a hydrate or solvate, a crystalline form,or a pharmaceutical composition in the manufacture of a medicament forthe treatment of an S1P1 receptor-associated disorder.

One aspect of the present invention pertains to the use of compounds ofthe present invention in the manufacture of a medicament for thetreatment of a S1P1 receptor associated disorder selected from the groupconsisting of: a disease or disorder mediated by lymphocytes, anautoimmune disease or disorder, an inflammatory disease or disorder,cancer, psoriasis, rheumatoid arthritis, Crohn's disease, transplantrejection, multiple sclerosis, systemic lupus erythematosus, ulcerativecolitis, type 1 diabetes and acne.

One aspect of the present invention pertains to the use of compounds ofthe present invention, a salt, a hydrate or solvate, a crystalline form,or a pharmaceutical composition in the manufacture of a medicament forthe treatment of a S1P1 receptor associated disorder selected from thegroup consisting of: a disease or disorder mediated by lymphocytes, anautoimmune disease or disorder, an inflammatory disease or disorder,cancer, psoriasis, rheumatoid arthritis, Crohn's disease, transplantrejection, multiple sclerosis, systemic lupus erythematosus, ulcerativecolitis, type I diabetes and acne.

One aspect of the present invention pertains to the use of compounds ofthe present invention in the manufacture of a medicament for thetreatment of a disease or disorder mediated by lymphocytes.

One aspect of the present invention pertains to the use of compounds ofthe present invention in the manufacture of a medicament for thetreatment of an autoimmune disease or disorder.

One aspect of the present invention pertains to the use of compounds ofthe present invention in the manufacture of a medicament for thetreatment of an inflammatory disease or disorder.

One aspect of the present invention pertains to the use of compounds ofthe present invention in the manufacture of a medicament for thetreatment of cancer.

One aspect of the present invention pertains to the use of compounds ofthe present invention in the manufacture of a medicament for thetreatment of an S1P1 receptor-associated disorder selected from thegroup consisting of psoriasis, rheumatoid arthritis, Crohn's disease,transplant rejection, multiple sclerosis, systemic lupus erythematosus,ulcerative colitis, type I diabetes and acne.

One aspect of the present invention pertains to the use of compounds ofthe present invention in the manufacture of a medicament for thetreatment of psoriasis.

One aspect of the present invention pertains to the use of compounds ofthe present invention in the manufacture of a medicament for thetreatment of rheumatoid arthritis.

One aspect of the present invention pertains to the use of compounds ofthe present invention in the manufacture of a medicament for thetreatment of Crohn's disease.

One aspect of the present invention pertains to the use of compounds ofthe present invention in the manufacture of a medicament for thetreatment of transplant rejection.

One aspect of the present invention pertains to the use of compounds ofthe present invention in the manufacture of a medicament for thetreatment of multiple sclerosis.

One aspect of the present invention pertains to the use of compounds ofthe present invention in the manufacture of a medicament for thetreatment of systemic lupus erythematosus.

One aspect of the present invention pertains to the use of compounds ofthe present invention in the manufacture of a medicament for thetreatment of ulcerative colitis.

One aspect of the present invention pertains to the use of compounds ofthe present invention in the manufacture of a medicament for thetreatment of type I diabetes.

One aspect of the present invention pertains to the use of compounds ofthe present invention in the manufacture of a medicament for thetreatment of acne.

One aspect of the present invention pertains to the use of compounds ofthe present invention in the manufacture of a medicament for thetreatment of a S1P1 receptor associated disorder wherein the S1P1receptor associated disorder is a microbial infection or disease or aviral infection or disease.

One aspect of the present invention pertains to the use of compounds ofthe present invention, a salt, a hydrate or solvate, a crystalline form,or a pharmaceutical composition in the manufacture of a medicament forthe treatment of a S1P1 receptor associated disorder wherein the S1P1receptor associated disorder is a microbial infection or disease or aviral infection or disease.

One aspect of the present invention pertains to the use of compounds ofthe present invention in the manufacture of a medicament for thetreatment of gastritis.

One aspect of the present invention pertains to the use of compounds ofthe present invention in the manufacture of a medicament for thetreatment of polymyositis.

One aspect of the present invention pertains to the use of compounds ofthe present invention in the manufacture of a medicament for thetreatment of thyroiditis.

One aspect of the present invention pertains to the use of compounds ofthe present invention in the manufacture of a medicament for thetreatment of vitiligo.

One aspect of the present invention pertains to the use of compounds ofthe present invention in the manufacture of a medicament for thetreatment of hepatitis.

One aspect of the present invention pertains to the use of compounds ofthe present invention in the manufacture of a medicament for thetreatment of biliary cirrhosis.

One aspect of the present invention pertains to compounds of the presentinvention for use in a method for the treatment of the human or animalbody by therapy.

One aspect of the present invention pertains to compounds of the presentinvention, a salt, a hydrate or solvate, a crystalline form, for use ina method for the treatment of the human or animal body by therapy.

One aspect of the present invention pertains to compounds of the presentinvention for use in a method for the treatment of an S1P1receptor-associated disorder.

One aspect of the present invention pertains to compounds of the presentinvention, a salt, a hydrate or solvate, a crystalline form, for use ina method for the treatment of an S1P1 receptor-associated disorder.

One aspect of the present invention pertains to compounds of the presentinvention for use in a method for the treatment of a S1P1 receptorassociated disorder selected from the group consisting of a disease ordisorder mediated by lymphocytes, an autoimmune disease or disorder, aninflammatory disease or disorder, cancer, psoriasis, rheumatoidarthritis, Crohn's disease, transplant rejection, multiple sclerosis,systemic lupus erythematosus, ulcerative colitis, type I diabetes andacne.

One aspect of the present invention pertains to compounds of the presentinvention, a salt, a hydrate or solvate, a crystalline form, for use ina method for the treatment of a S1P1 receptor associated disorderselected from the group consisting of: a disease or disorder mediated bylymphocytes, an autoimmune disease or disorder, an inflammatory diseaseor disorder, cancer, psoriasis, rheumatoid arthritis, Crohn's disease,transplant rejection, multiple sclerosis, systemic lupus erythematosus,ulcerative colitis, type I diabetes and acne.

One aspect of the present invention pertains to compounds of the presentinvention for use in a method for the treatment of a disease or disordermediated by lymphocytes.

One aspect of the present invention pertains to compounds of the presentinvention for use in a method for the treatment of an autoimmune diseaseor disorder.

One aspect of the present invention pertains to compounds of the presentinvention for use in a method for the treatment of an inflammatorydisease or disorder.

One aspect of the present invention pertains to compounds of the presentinvention for use in a method for the treatment of cancer.

One aspect of the present invention pertains to compounds of the presentinvention for use in a method for the treatment of an S1P1receptor-associated disorder selected from the group consisting ofpsoriasis, rheumatoid arthritis, Crohn's disease, transplant rejection,multiple sclerosis, systemic lupus erythematosus, ulcerative colitis,type I diabetes and acne.

One aspect of the present invention pertains to compounds of the presentinvention for use in a method for the treatment of psoriasis.

One aspect of the present invention pertains to compounds of the presentinvention for use in a method for the treatment of rheumatoid arthritis.

One aspect of the present invention pertains to compounds of the presentinvention for use in a method for the treatment of Crohn's disease.

One aspect of the present invention pertains to compounds of the presentinvention for use in a method for the treatment of transplant rejection.

One aspect of the present invention pertains to compounds of the presentinvention for use in a method for the treatment of multiple sclerosis.

One aspect of the present invention pertains to compounds of the presentinvention for use in a method for the treatment of systemic lupuserythematosus.

One aspect of the present invention pertains to compounds of the presentinvention for use in a method for the treatment of ulcerative colitis.

One aspect of the present invention pertains to compounds of the presentinvention for use in a method for the treatment of type I diabetes.

One aspect of the present invention pertains to compounds of the presentinvention for use in a method for the treatment of acne.

One aspect of the present invention pertains to compounds of the presentinvention for use in a method for the treatment of a S1P1 receptorassociated disorder wherein the S1P1 receptor associated disorder is amicrobial infection or disease or a viral infection or disease.

One aspect of the present invention pertains to compounds of the presentinvention, a salt, a hydrate or solvate, a crystalline form, for use ina method for the treatment of a S1P1 receptor associated disorderwherein the S1P1 receptor associated disorder is a microbial infectionor disease or a viral infection or disease.

One aspect of the present invention pertains to compounds of the presentinvention for use in a method for the treatment of gastritis.

One aspect of the present invention pertains to compounds of the presentinvention for use in a method for the treatment of polymyositis.

One aspect of the present invention pertains to compounds of the presentinvention for use in a method for the treatment of thyroiditis.

One aspect of the present invention pertains to compounds of the presentinvention for use in a method for the treatment of vitiligo.

One aspect of the present invention pertains to compounds of the presentinvention for use in a method for the treatment of hepatitis.

One aspect of the present invention pertains to compounds of the presentinvention for use in a method for the treatment of biliary cirrhosis.

One aspect of the present invention pertains to processes for preparinga composition comprising admixing a compound of the present inventionand a pharmaceutically acceptable carrier.

One aspect of the present invention pertains to processes for preparinga composition comprising admixing a compound of the present invention, asalt, a hydrate or solvate, a crystalline form and a pharmaceuticallyacceptable carrier.

These and other aspects of the invention disclosed herein will be setforth in greater detail as the patent disclosure proceeds.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 shows the results of an experiment which measured the ability ofCompound 7 to lower the absolute count of peripheral lymphocytes in micecompared to vehicle.

FIG. 2 shows the results of an experiment which measured the ability ofCompound 5 to lower the absolute count of peripheral lymphocytes in micecompared to vehicle.

FIG. 3 shows a general synthetic scheme for the preparation of1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetic acid derivatives, viacoupling of the aryl methyl halides or alcohols with ethyl2-(7-hydroxy-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetate.Subsequent hydrolysis of the ester functionality affords compounds ofFormula (Ia) wherein “m” is 1 and “n” is 1.

FIG. 4 shows a general synthetic scheme for the preparation of anhalogenated 1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetic acidintermediate, via coupling of the aryl methyl halides with ethyl2-(7-hydroxy-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetate.Subsequent functionalization at the aromatic halogen and hydrolysis ofthe ester functionality afford compounds of Formula (Ia) wherein “m” is1 and “n” is 1.

FIG. 5 shows a general synthetic scheme for the preparation of alcoholintermediates used in the preparation of compounds of Formula (Ia). Thesynthetic scheme shows the functionalization at the aromatic halogen bymetal-catalyzed coupling, followed by conversion of the hydroxyl groupto a triflate moiety. Subsequent replacement of the triflate with avariety of functional groups by metal-catalyzed coupling reactions andreduction of the ester moiety afforded alcohol intermediates.

FIG. 6 shows a general synthetic scheme for the preparation of bromideintermediates used in the preparation of compounds of Formula (Ia). Thesynthetic scheme shows the functionalization at the aromatic halogen bymetal-catalyzed coupling or nucleophilic displacement. Subsequentbromination of the methyl group affords bromide intermediates.

FIG. 7 shows the results of an experiment which measured the ability ofthe 2^(nd) enantiomer of compound 12 (isolated after resolution ofcompound 12 by HPLC with a retention time of 13.9 min per the conditionsreported in Example 1.29) to lower the absolute count of peripherallymphocytes in mice compared to vehicle.

FIG. 8 shows the results of an experiment which measured the ability ofthe 2^(nd) enantiomer of compound 12 (isolated after resolution ofcompound 12 by HPLC with a retention time of 13.9 min per the conditionsreported in Example 1.29) to lower the absolute count of peripherallymphocytes in rats compared to vehicle.

FIG. 9 shows the results of an experiment which measured the ability ofthree different doses of the 2^(nd) enantiomer of compound 12 (isolatedafter resolution of compound 12 by HPLC with a retention time of 13.9min per the conditions reported in Example 1.29) to reduce the meanankle diameter in rats compared to vehicle.

FIG. 10 shows the results of an experiment which measured the ability ofthree different doses of the 2^(nd) enantiomer of compound 12 (isolatedafter resolution of compound 12 by HPLC, with a retention time of 119min per the conditions reported in Example 1.29) to have efficacy inexperimental autoimmune encephalomyelitis (EAE) compared to vehicle.

FIG. 11 shows the results of an experiment wherein no or substantiallyno reduction of heart rate was exhibited in response to the treatment ofrats with the 2^(nd) enantiomer of compound 12 (isolated afterresolution of compound 12 by HPLC with a retention time of 13.9 min perthe conditions reported in Example 1.29) in comparison with vehicle.

FIG. 12 depicts a powder X-ray diffraction (PXRD) pattern for a crystalform of the 2^(nd) enantiomer of compound 12 (isolated after resolutionof compound 12 by HPLC with a retention time of 13.9 min per theconditions reported in Example 1.29).

FIG. 13 depicts a differential scanning calorimetry (DSC) thermogram anda thermogravimetric analysis (TGA) thermogram for the 2^(nd) enantiomerof compound 12 (isolated after resolution of compound 12 by HPLC, with aretention time of 13.9 min per the conditions reported in Example 1.29).

FIG. 14 depicts a moisture sorption analysis for the 2^(nd) enantiomerof compound 12 (isolated after resolution of compound 12 by HPLC, with aretention time of 13.9 min per the conditions reported in Example 1.29).

FIG. 15 depicts a powder X-ray diffraction (PXRD) pattern for a crystalform of the Ca salt of the 2^(nd) enantiomer of compound 12 (asdescribed in Example 1.32).

FIG. 16 depicts a differential scanning calorimetry (DSC) thermogram forthe Ca salt of the 2^(nd) enantiomer of compound 12 (as described inExample 1.32).

FIG. 17 depicts a thermogravimetric analysis (TGA) thermogram for the Casalt of the 2^(nd) enantiomer of compound 12 (as described in Example1.32).

FIG. 18 depicts a powder X-ray diffraction (PXRD) pattern for a crystalform of the D-Lysine salt of the 1^(st) enantiomer of compound 12 (asdescribed in Example 1.34).

FIG. 19 depicts a differential scanning calorimetry (DSC) thermogram forthe D-Lysine salt of the 1^(st) enantiomer of compound 12 (as describedin Example 1.34).

FIG. 20 depicts a thermogravimetric analysis (TGA) thermogram for theD-Lysine salt of the 1^(st) enantiomer of compound 12 (as described inExample 1.34).

FIG. 21 depicts a view of a molecule that appears to be the 1^(st)enantiomer of compound 12 (isolated after resolution of compound 12 byHPLC with a retention time of 9.1 min per the conditions reported inExample 1.29).

FIG. 22 shows a general synthetic scheme for the preparation of1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetic acid derivatives, viathe Fisher indole synthesis. Subsequent hydrolysis and decarboxylationaffords compounds of Formula (Ia) wherein “m” is 1 and “n” is 1.

FIG. 23 depicts a powder X-ray diffraction (PXRD) pattern for a crystalform of the L-Arginine salt of the 2^(nd) enantiomer of compound 12 (asdescribed in Example 1.33).

FIG. 24 depicts a differential scanning calorimetry (DSC) thermogram forthe L-Arginine salt of the 2^(nd) enantiomer of compound 12 (asdescribed in Example 1.33).

FIG. 25 depicts a thermogravimetric analysis (TGA) thermogram for theL-Arginine salt of the 2^(nd) enantiomer of compound 12 (as described inExample 1.33).

FIG. 26 depicts a moisture sorption analysis for the L-Arginine salt ofthe 2^(nd) enantiomer of compound 12 (as described in Example 1.33).

DETAILED DESCRIPTION OF THE INVENTION Definitions

For clarity and consistency, the following definitions will be usedthroughout this patent document.

The term “agonist” is intended to mean a moiety that interacts with andactivates a G-protein-coupled receptor, such as the S1P1 receptor, suchas can thereby initiate a physiological or pharmacological responsecharacteristic of that receptor. For example, an agonist activities anintracellular response upon binding to the receptor, or enhances GTPbinding to a membrane. In certain embodiments, an agonist of theinvention is an S1P1 receptor agonist that is capable of facilitatingsustained S1P1 receptor internalization (see e.g., Matloubian et al.,Nature, 427, 355, 2004).

The term “antagonist” is intended to mean a moiety that competitivelybinds to the receptor at the same site as an agonist (for example, theendogenous ligand), but which does not activate the intracellularresponse initiated by the active form of the receptor and can therebyinhibit the intracellular responses by an agonist or partial agonist. Anantagonist does not diminish the baseline intracellular response in theabsence of an agonist or partial agonist.

The term “hydrate” as used herein means a compound of the invention or asalt thereof, that further includes a stoichiometric ornon-stoichiometric amount of water bound by non-covalent intermolecularforces.

The term “solvate” as used herein means a compound of the invention or asalt, thereof, that further includes a stoichiometric ornon-stoichiometric amount of a solvent bound by non-covalentintermolecular forces. Preferred solvents are volatile, non-toxic,and/or acceptable for administration to humans in trace amounts.

The term “in need of treatment” and the term “in need thereof” whenreferring to treatment are used interchangeably to mean a judgment madeby a caregiver (e.g. physician, nurse, nurse practitioner, etc. in thecase of humans; veterinarian in the case of animals, including non-humanmammals) that an individual or animal requires or will benefit fromtreatment. This judgment is made based on a variety of factors that arein the realm of a caregiver's expertise, but that includes the knowledgethat the individual or animal is ill, or will become ill, as the resultof a disease, condition or disorder that is treatable by the compoundsof the invention. Accordingly, the compounds of the invention can beused in a protective or preventive manner; or compounds of the inventioncan be used to alleviate, inhibit or ameliorate the disease, conditionor disorder.

The term “individual” is intended to mean any animal, including mammals,preferably mice, rats, other rodents, rabbits, dogs, cats, swine,cattle, sheep, horses, or primates and most preferably humans.

The term “inverse agonist” is intended to mean a moiety that binds tothe endogenous form of the receptor or to the constitutively activatedform of the receptor and which inhibits the baseline intracellularresponse initiated by the active form of the receptor below the normalbase level of activity which is observed in the absence of an agonist orpartial agonist, or decreases GTP binding to a membrane. In someembodiments, the baseline intracellular response is inhibited in thepresence of the inverse agonist by at least 30%. In some embodiments,the baseline intracellular response is inhibited in the presence of theinverse agonist by at least 50%. In some embodiments, the baselineintracellular response is inhibited in the presence of the inverseagonist by at least 75%, as compared with the baseline response in theabsence of the inverse agonist.

The term “modulate or modulating” is intended to mean an increase ordecrease in the amount, quality, response or effect of a particularactivity, function or molecule.

The term “pharmaceutical composition” is intended to mean a compositioncomprising at least one active ingredient; including but not limited to,salts, solvates and hydrates of compounds of the present invention,whereby the composition is amenable to investigation for a specified,efficacious outcome in a mammal (for example, without limitation, ahuman). Those of ordinary skill in the art will understand andappreciate the techniques appropriate for determining whether an activeingredient has a desired efficacious outcome based upon the needs of theartisan.

The term “therapeutically effective amount” is intended to mean theamount of active compound or pharmaceutical agent that elicits thebiological or medicinal response in a tissue, system, animal, individualor human that is being sought by a researcher, veterinarian, medicaldoctor or other clinician, caregiver or by an individual, which includesone or more of the following:

(1) Preventing the disease, for example, preventing a disease, conditionor disorder in an individual that may be predisposed to the disease,condition or disorder but does not yet experience or display thepathology or symptomatology of the disease;

(2) Inhibiting the disease, for example, inhibiting a disease, conditionor disorder in an individual that is experiencing or displaying thepathology or symptomatology of the disease, condition or disorder (i.e.,arresting further development of the pathology and/or symptomatology);and

(3) Ameliorating the disease, for example, ameliorating a disease,condition or disorder in an individual that is experiencing ordisplaying the pathology or symptomatology of the disease, condition ordisorder (i.e., reversing the pathology and/or symptomatology).

Chemical Group, Moiety or Radical

The term “C₁-C₆ alkoxy” is intended to mean a C₁-C₆ alkyl radical, asdefined herein, attached directly to an oxygen atom. Some embodimentsare 1 to 5 carbons, some embodiments are 1 to 4 carbons, someembodiments are 1 to 3 carbons and some embodiments are 1 or 2 carbons.Examples include methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy,tert-butoxy, isobutoxy, sec-butoxy and the like.

The term “C₁-C₆ alkyl” is intended to mean a straight or branched carbonradical containing 1 to 6 carbons. Some embodiments are 1 to 5 carbons,some embodiments are 1 to 4 carbons, some embodiments are 1 to 3 carbonsand some embodiments are 1 or 2 carbons. Examples of an alkyl include,but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl,sec-butyl, isobutyl, tert-butyl, pentyl, isopentyl, tert-pentyl,neo-pentyl, 1-methylbutyl [i.e., —CH(CH₃)CH₂CH₂CH₃], 2-methylbutyl[i.e., —CH₂CH(CH₃)CH₂CH₃], n-hexyl and the like.

The term “C₁-C₆ alkylamino” is intended to mean one alkyl radicalattached to an —NH-radical wherein the alkyl radical has the samemeaning as described herein. Some examples include, but are not limitedto, methylamino, ethylamino, n-propylamino, isopropylamino,n-butylamino, sec-butylamino, isobutylamino, tert-butylamino and thelike.

The term “C₁-C₆ alkylsulfonyl” is intended to mean a C₁-C₆ alkyl radicalattached to the sulfur of a sulfone radical having the formula: —S(O)₂—wherein the alkyl radical has the same definition as described herein.Examples include, but are not limited to, methylsulfonyl, ethylsulfonyl,n-propylsulfonyl, iso-propylsulfonyl, n-butylsulfonyl,sec-butylsulfonyl, iso-butylsulfonyl, tert-butylsulfonyl and the like.

The term “carboxamide” is intended to mean the group —CONH₂.

The term “carboxy” or “carboxyl” is intended to mean the group —CO₂H;also referred to as a carboxylic acid group.

The term “cyano” is intended to mean the group —CN.

The term “C₃-C₇ cycloalkoxy” is intended to mean a saturated ringradical containing 3 to 7 carbons directly bonded to an oxygen atom.Some examples include cyclopropyl-O—, cyclobutyl-O—, cyclopentyl-O—,cyclohexyl-O— and the like.

The term “C₃-C₇ cycloalkyl” is intended to mean a saturated ring radicalcontaining 3 to 7 carbons. Some embodiments contain 3 to 6 carbons. Someembodiments contain 3 to 5 carbons. Some embodiments contain 5 to 7carbons. Some embodiments contain 3 to 4 carbons. Examples includecyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl and thelike.

The term “C₁-C₆ haloalkoxy” is intended to mean a C₁-C₆ haloalkyl, asdefined herein, which is directly attached to an oxygen atom. Examplesinclude, but are not limited to, difluoromethoxy, trifluoromethoxy,2,2,2-trifluoroethoxy, pentafluoroethoxy and the like.

The term “C₁-C₆ haloalkyl” is intended to mean an C₁-C₆ alkyl group,defined herein, wherein the alkyl is substituted with between onehalogen up to fully substituted wherein a fully substituted C₁-C₆haloalkyl can be represented by the formula C_(n)L_(2n+1) wherein L is ahalogen and “n” is 1, 2, 3, 4, 5 or 6. When more than one halogen ispresent, the halogens may be the same or different and selected from thegroup consisting of fluoro, chloro, bromo or iodo, preferably fluoro.Some embodiments are 1 to 5 carbons, some embodiments are 1 to 4carbons, some embodiments are 1 to 3 carbons and some embodiments are 1or 2 carbons. Examples of haloalkyl groups include, but are not limitedto, fluoromethyl, difluoromethyl, trifluoromethyl, chlorodifluoromethyl,2,2,2-trifluoroethyl, pentafluoroethyl and the like.

The term “halogen” or “halo” is intended to mean a fluoro, chloro, bromoor iodo group.

The term “heteroaryl” is intended to mean an aromatic ring systemcontaining 5 to 14 aromatic ring atoms that may be a single ring, twofused rings or three fused rings, wherein at least one aromatic ringatom is a heteroatom selected from, for example, but not limited to, thegroup consisting of O, S and N wherein the N can be optionallysubstituted with H, C₁-C₄ acyl or C₁-C₄ alkyl. Some embodiments contain5 to 6 ring atoms, for example, furanyl, thienyl, pyrrolyl, imidazolyl,oxazolyl, thiazolyl, isoxazolyl, pyrazolyl, isothiazolyl, oxadiazolyl,triazolyl, thiadiazolyl, pyridinyl, pyrazinyl, pyrimidinyl, pyridazinyland triazinyl and the like. Some embodiments contain 8 to 14 ring atoms,for example, quinolizinyl, quinolinyl, isoquinolinyl, cinnolinyl,phthalazinyl, quinazolinyl, quinoxalinyl, triazinyl, indolyl,isoindolyl, indazolyl, indolizinyl, purinyl, naphthyridinyl, pteridinyl,carbazolyl, acridinyl, phenazinyl, phenothiazinyl, phenoxazinyl,benzoxazolyl, benzothiazolyl, 1H-benzimidazolyl, imidazopyridinyl,benzothienyl, benzofuranyl, and isobenzofuran and the like.

The term “heterocyclic” or “heterocyclyl” is intended to mean anon-aromatic ring containing 3 to 8 ring atoms wherein one, two or threering atoms are heteroatoms selected from, for example, the groupconsisting of O, S, S(═O), S(═O)₂ and NH, wherein the N is optionallysubstituted as described herein. In some embodiments, the nitrogen isoptionally substituted with C₁-C₄ acyl or C₁-C₄ alkyl. In someembodiments, ring carbon atoms are optionally substituted with oxo thusforming a carbonyl group. In some embodiments, ring sulfur atoms areoptionally substituted with oxo atoms thus forming a thiocarbonyl group.The heterocyclic group can be attached/bonded to any available ringatom, for example, ring carbon, ring nitrogen and the like. In someembodiments the heterocyclic group is a 3-, 4-, 5-, 6- or 7-memberedring. Examples of a heterocyclic group include, but are not limited to,aziridin-1-yl, aziridin-2-yl, azetidin-1-yl, azetidin-2-yl,azetidin-3-yl, piperidin-1-yl, piperidin-2-yl, piperidin-3-yl,piperidin-4-yl, morpholin-2-yl, morpholin-3-yl, morpholin-4-yl,piperzin-1-yl, piperzin-2-yl, piperzin-3-yl, piperzin-4-yl,pyrrolidin-1-yl, pyrrolidin-2-yl, pyrrolidin-3-yl, [1,3]-dioxolan-2-yl,thiomorpholin-4-yl, [1,4]oxazepan-4-yl, 1,1-dioxothiomorpholin-4-yl,azepan-1-yl, azepan-2-yl, azepan-3-yl, azepan-4-yl,tetrahydrofuran-2-yl, tetrahydrofuran-3-yl and the like.

Compounds of the Invention

One aspect of the present invention pertains to certain compounds ofFormula (Ia) and pharmaceutically acceptable salts, solvates andhydrates thereof:

wherein:

m, n, R², R³, Y and Z have the same definitions as described herein,supra and infra.

It is understood that the present invention embraces compounds, solvatesand/or hydrates of compounds, pharmaceutically acceptable salts ofcompounds, and solvates and/or hydrates of pharmaceutically acceptablesalts of compounds, wherein the compounds are as described herein.

It is appreciated that certain features of the invention, which are, forclarity, described in the context of separate embodiments, may also beprovided in combination in a single embodiment. Conversely, variousfeatures of the invention, which are, for brevity, described in thecontext of a single embodiment, may also be provided separately or inany suitable subcombination. All combinations of the embodimentspertaining to the chemical groups represented by the variables (e.g.,in, n, R¹, R², R³, Y and Z) contained within the generic chemicalformulae described herein, for example, (Ia), (Ic), (Ie), (Ig), (Ii),(Ik), (Im) are specifically embraced by the present invention just as ifeach and every combination was individually explicitly recited, to theextent that such combinations embrace stable compounds (i.e., compoundsthat can be isolated, characterized and tested for biological activity).In addition, all subcombinations of the chemical groups listed in theembodiments describing such variables, as well as all subcombinations ofuses and medical indications described herein, are also specificallyembraced by the present invention just as if each and everysubcombination of chemical groups and subcombination of uses and medicalindications was individually and explicitly recited herein.

As used herein, “substituted” indicates that at least one hydrogen atomof the chemical group is replaced by a non-hydrogen substituent orgroup. The non-hydrogen substituent or group can be monovalent ordivalent. When the substituent or group is divalent, then it isunderstood that this group is further substituted with anothersubstituent or group. When a chemical group herein is “substituted” itmay have up to the full valence of substitution, for example, a methylgroup can be substituted by 1, 2, or 3 substituents, a methylene groupcan be substituted by 1 or 2 substituents, a phenyl group can besubstituted by 1, 2, 3, 4, or 5 substituents, a naphthyl group can besubstituted by 1, 2, 3, 4, 5, 6, or 7 substituents and the like.Likewise, “substituted with one or more substituents” refers to thesubstitution of a group with one substituent up to the total number ofsubstituents physically allowed by the group. Further, when a group issubstituted with more than one substituent, the substituents can beidentical or they can be different.

Compounds of the invention also include tautomeric forms, such asketo-enol tautomers and the like. Tautomeric forms can be in equilibriumor sterically locked into one form by appropriate substitution. It isunderstood that the various tautomeric forms are within the scope of thecompounds of the present invention.

Compounds of the invention also include all isotopes of atoms occurringin the intermediates and/or final compounds. Isotopes include thoseatoms having the same atomic number but different mass numbers. Forexample, isotopes of hydrogen include deuterium and tritium.

It is understood and appreciated that compounds of Formula (Ia) andformulae related thereto may have one or more chiral centers andtherefore can exist as enantiomers and/or diastereomers. The inventionis understood to extend to and embrace all such enantiomers,diastereomers and mixtures thereof, including but not limited toracemates. It is understood that Formula (Ia) and formulae usedthroughout this disclosure are intended to represent all individualenantiomers and mixtures thereof, unless stated or shown otherwise.

The Variable “n”

In some embodiments, n is 1.

In some embodiments, compounds of the present invention are representedby Formula (Ic) as illustrated below:

wherein each variable in Formula (Ic) has the same meaning as describedherein, supra and infra.

In some embodiments, n is 2.

In some embodiments, compounds of the present invention are representedby Formula (Ie) as illustrated below:

wherein each variable in Formula (Ie) has the same meaning as describedherein, supra and infra.The Variable “m”

In some embodiments, m is 1.

In some embodiments, compounds of the present invention are representedby Formula (Ig) as illustrated below:

wherein each variable in Formula (Ig) has the same meaning as describedherein, supra and infra.

In some embodiments, m is 2.

In some embodiments, compounds of the present invention are representedby Formula (Ii) as illustrated below:

wherein each variable in Formula (Ii) has the same meaning as describedherein, supra and infra.

The Variables Y and Z

In some embodiments, Y is N or CR¹ and Z is N or CR⁴.

In some embodiments, Y is N and Z is N.

In some embodiments, Y is N and Z is CR⁴.

In some embodiments, Y is CR¹ and Z is N.

In some embodiments, Y is CR¹ and Z is CR⁴.

In some embodiments, Y is N.

In some embodiments, Y is CR¹.

In some embodiments, Z is N.

In some embodiments, Z is CR⁴.

The Group R¹

In some embodiments, R¹ is selected from the group consisting of H,C₁-C₆ alkoxy, C₆ alkyl, C₁-C₆ alkylamino, C₁-C₆ alkylsulfonyl, C₁-C₆alkylthio, carboxamide, cyano, C₃-C₇ cycloalkoxy, C₃-C₇ cycloalkyl,C₁-C₆ haloalkoxy, C₁-C₆ haloalkyl, halogen, heteroaryl and heterocyclyl,wherein said C₁-C₆ alkyl and C₁-C₆ alkoxy are each optionallysubstituted with one or two substituents selected from C₃-C₇ cycloalkyland halogen.

In some embodiments, R¹ is selected from the group consisting of H,C₁-C₆ alkoxy, C₁-C₆ alkyl, C₁-C₆ alkylamino, C₁-C₆ alkylsulfonyl, C₁-C₆alkylthio, carboxamide, cyano, C₃-C₇ cycloalkoxy, C₃-C₇ cycloalkyl,C₁-C₆ haloalkoxy, C₁-C₆ haloalkyl, halogen, heteroaryl and heterocyclyl,wherein said C₁-C₆ alkyl and C₁-C₆ alkoxy are each optionallysubstituted with one C₃-C₇ cycloalkyl group.

In some embodiments, R¹ is H or C₁-C₆ haloalkyl.

In some embodiments, R¹ is H.

In some embodiments, R¹ is trifluoromethyl.

The Group R²

In some embodiments, R² is selected from the group consisting of H,C₁-C₆ alkoxy, C₁-C₆ alkyl, C₁-C₆ alkylamino, C₁-C₆ alkylsulfonyl, C₁-C₆alkylthio, carboxamide, cyano, C₃-C₇ cycloalkoxy, C₃-C₇ cycloalkyl,C₁-C₆ haloalkoxy, C₁-C₆ haloalkyl, halogen, heteroaryl and heterocyclyl,wherein said C₁-C₆ alkyl and C₁-C₆ alkoxy are each optionallysubstituted with one or two substituents selected from C₃-C₇ cycloalkyland halogen.

In some embodiments, R² is selected from the group consisting of H,C₁-C₆ alkoxy, C₁-C₆ alkyl, C₁-C₆ alkylamino, C₁-C₆ alkylsulfonyl, C₁-C₆alkylthio, carboxamide, cyano, C₃-C₇ cycloalkoxy, C₃-C₇ cycloalkyl,C₁-C₆ haloalkoxy, C₁-C₆ haloalkyl, halogen, heteroaryl and heterocyclyl,wherein said C₁-C₆ alkyl and C₁-C₆ alkoxy are each optionallysubstituted with one C₃-C₇ cycloalkyl group.

In some embodiments, R² is selected from the group consisting of H,cyano, C₁-C₆ haloalkoxy and C₁-C₆ haloalkyl.

In some embodiments, R² is selected from the group consisting of H,cyano, trifluoromethoxy and trifluoromethyl.

In some embodiments, R² is H.

In some embodiments, R² is cyano.

In some embodiments, R² is trifluoromethoxy.

In some embodiments, R² is trifluoromethyl.

The Group R³

In some embodiments, R³ is selected from the group consisting of H,C₁-C₆ alkoxy, C₁-C₆ alkyl, C₁-C₆ alkylamino, C₁-C₆ alkylsulfonyl, C₁-C₆alkylthio, carboxamide, cyano, C₃-C₇ cycloalkoxy, C₃-C₇ cycloalkyl,C₁-C₆ haloalkoxy, C₁-C₆ haloalkyl, halogen, heteroaryl and heterocyclyl,wherein said C₁-C₆ alkyl and C₁-C₆ alkoxy are each optionallysubstituted with one or two substituents selected from C₃-C₇ cycloalkyland halogen.

In some embodiments, R³ is selected from the group consisting of H,C₁-C₆ alkoxy, C₁-C₆ alkyl, C₁-C₆ alkylamino, C₁-C₆ alkylsulfonyl,carboxamide, cyano, C₃-C₇ cycloalkoxy, C₃-C₇ cycloalkyl, C₁-C₆haloalkoxy, C₁-C₆ haloalkyl, halogen, heteroaryl and heterocyclyl,wherein said C₁-C₆ alkyl and C₁-C₆ alkoxy are each optionallysubstituted with one or two substituents selected from C₃-C₇ cycloalkyland halogen

In some embodiments, R³ is selected from the group consisting of H,C₁-C₆ alkoxy, C₁-C₆ alkyl, C₁-C₆ alkylamino, C₁-C₆ alkylsulfonyl, C₁-C₆alkylthio, carboxamide, cyano, C₃-C₇ cycloalkoxy, C₃-C₇ cycloalkyl,C₁-C₆ haloalkoxy, C₁-C₆ haloalkyl, halogen, heteroaryl and heterocyclyl,wherein said C₁-C₆ alkyl and C₁-C₆ alkoxy are each optionallysubstituted with one C₃-C₇ cycloalkyl group.

In some embodiments, R³ is selected from the group consisting of H,C₁-C₆ alkoxy, C₁-C₆ alkyl, C₃-C₇ cycloalkyl, C₁-C₆ haloalkoxy, C₁-C₆haloalkyl, halogen and heterocyclyl.

In some embodiments, R³ is selected from the group consisting ofselected from the group consisting of H, carboxamide, chloro, cyano,cyclobutyl, cyclohexyl, cyclopentyl, cyclopentyloxy, cyclopropyl,cyclopropylmethoxy, cyclohexylmethyl, 3,3-difluoropyrrolidin-1-yl,ethylamino, isobutyl, isopropoxy, methylsulfonyl, neopentyl, propyl,pyrrolidin-1-yl, 1,2,3-thiadiazol-4-yl, trifluoromethoxy andtrifluoromethyl.

In some embodiments, R³ is selected from the group consisting of H,chloro, cyclobutyl, cyclohexyl, cyclopentyl, cyclopropyl,3,3-difluoropyrrolidin-1-yl, isobutyl, isopropoxy, neopentyl, propyl,pyrrolidin-1-yl, trifluoromethoxy and trifluoromethyl.

In some embodiments, R³ is H.

In some embodiments, R³ is chloro.

In some embodiments, R³ is cyclobutyl.

In some embodiments, R³ is cyclohexyl.

In some embodiments, R³ is cyclopentyl.

In some embodiments, R³ is cyclopropyl.

In some embodiments, R³ is 3,3-difluoropyrrolidin-1-yl.

In some embodiments, R³ is isobutyl.

In some embodiments, R³ is isopropoxy.

In some embodiments, R³ is neopentyl.

In some embodiments, R³ is propyl.

In some embodiments, R³ is pyrrolidin-1-yl.

In some embodiments, R³ is trifluoromethoxy.

In some embodiments, R³ is trifluoromethyl.

In some embodiments, R³ is carboxamide.

In some embodiments, R³ is cyano.

In some embodiments, R³ is cyclopentyloxy.

In some embodiments, R³ is cyclopropylmethoxy.

In some embodiments, R³ is cyclohexylmethyl.

In some embodiments, R³ is ethylamino.

In some embodiments, R³ is methylsulfonyl.

In some embodiments, R³ is 1,2,3-thiadiazol-4-yl.

The Group R⁴

In some embodiments, R⁴ is selected from the group consisting of H,C₁-C₆ alkoxy, C₁-C₆ alkyl, C₁-C₆ alkylamino, C₁-C₆ alkylsulfonyl, C₁-C₆alkylthio, carboxamide, cyano, C₃-C₇ cycloalkoxy, C₃-C₇ cycloalkyl,C₁-C₆ haloalkoxy, C₁-C₆ haloalkyl, halogen, heteroaryl and heterocyclyl,wherein said C₁-C₆ alkyl and C₁-C₆ alkoxy are each optionallysubstituted with one or two substituents selected from C₃-C₇ cycloalkyland halogen.

In some embodiments, R⁴ is selected from the group consisting of H,C₁-C₆ alkoxy, C₁-C₆ alkyl, C₁-C₆ alkylamino, C₁-C₆ alkylsulfonyl, C₁-C₆alkylthio, carboxamide, cyano, C₃-C₇ cycloalkoxy, C₃-C₇ cycloalkyl,C₁-C₆ haloalkoxy, C₁-C₆ haloalkyl, halogen, heteroaryl and heterocyclyl,wherein said C₁-C₆ alkyl and C₁-C₆ alkoxy are each optionallysubstituted with one C₃-C₇ cycloalkyl group.

In some embodiments, R⁴ is selected from the group consisting of H,cyano, C₁-C₆ haloalkoxy and C₁-C₆ haloalkyl.

In some embodiments, R⁴ is selected from the group consisting of H,cyano and C₁-C₆ haloalkyl.

In some embodiments, R⁴ is selected from the group consisting of H,cyano, trifluoromethoxy and trifluoromethyl.

In some embodiments, R⁴ is selected from the group consisting of H,cyano and trifluoromethyl.

In some embodiments, R⁴ is H.

In some embodiments, R⁴ is cyano.

In some embodiments, R⁴ is trifluoromethyl.

In some embodiments, R⁴ is trifluoromethoxy.

Certain Combinations

Some embodiments of the present invention pertain to compounds selectedfrom compounds of Formula (Ia) and pharmaceutically acceptable salts,solvates and hydrates thereof; wherein:

m is 1 or 2;

n is 1 or 2;

Y is N or CR¹;

Z is N or CR⁴;

R¹ is H or C₁-C₆ haloalkyl;

R² is selected from the group consisting of H, cyano, C₁-C₆ haloalkoxyand C₁-C₆ haloalkyl;

R³ is selected from the group consisting of H, C₁-C₆ alkoxy, C₁-C₆alkyl, C₃-C₇ cycloalkyl, C₁-C₆ haloalkoxy, C₁-C₆ haloalkyl, halogen andheterocyclyl; and

R⁴ is selected from the group consisting of H, cyano and C₁-C₆haloalkyl.

Some embodiments of the present invention pertain to compounds selectedfrom compounds of Formula (Ia) and pharmaceutically acceptable salts,solvates and hydrates thereof, wherein:

m is 1 or 2;

n is 1 or 2;

Y is N or CR¹;

Z is N or CR⁴;

R¹ is H or trifluoromethyl;

R² is selected from the group consisting of H, cyano, trifluoromethoxyand trifluoromethyl;

R³ is selected from the group consisting of H, chloro, cyclobutyl,cyclohexyl, cyclopentyl, cyclopropyl, 3,3-difluoropyrrolidin-1-yl,isobutyl, isopropoxy, neopentyl, propyl, pyrrolidin-1-yl,trifluoromethoxy and trifluoromethyl; and

R⁴ is selected from the group consisting of H, cyano andtrifluoromethyl.

Some embodiments of the present invention pertain to compounds selectedfrom compounds of Formula (Ik) and pharmaceutically acceptable salts,solvates and hydrates thereof:

-   -   wherein:    -   Y is N or CR¹;    -   Z is N or CR⁴;    -   R¹ is H or C₁-C₆ haloalkyl;    -   R² is selected from the group consisting of H, cyano, C₁-C₆        haloalkoxy and C₁-C₆ haloalkyl;    -   R³ is selected from the group consisting of H, C₁-C₆ alkoxy,        C₁-C₆ alkyl, C₁-C₆ alkylamino, C₁-C₆ alkylsulfonyl, carboxamide,        cyano, C₃-C₇ cycloalkoxy, C₃-C₇ cycloalkyl, C₁-C₆ haloalkoxy,        C₁-C₆ haloalkyl, halogen, heteroaryl and heterocyclyl, wherein        said C₁-C₆ alkyl and C₁-C₆ alkoxy are each optionally        substituted with one or two substituents selected from C₃-C₇        cycloalkyl and halogen; and    -   R⁴ is selected from the group consisting of H, cyano, C₁-C₆        haloalkoxy and C₁-C₆ haloalkyl.

Some embodiments of the present invention pertain to compounds selectedfrom compounds of Formula (Ik) and pharmaceutically acceptable salts,solvates and hydrates thereof:

wherein:

Y is N or CR¹;

Z is N or CR⁴;

R¹ is H or C₁-C₆ haloalkyl;

R² is selected from the group consisting of H, cyano, C₁-C₆ haloalkoxyand C₁-C₆ haloalkyl;

R³ is selected from the group consisting of H, C₁-C₆ alkoxy, C₁-C₆alkyl, C₃-C₇ cycloalkyl, C₁-C₆ haloalkoxy, C₁-C₆ haloalkyl, halogen andheterocyclyl; and

R⁴ is selected from the group consisting of H, cyano and C₁-C₆haloalkyl.

Some embodiments of the present invention pertain to compounds selectedfrom compounds of Formula (Ik) and pharmaceutically acceptable salts,solvates and hydrates thereof:

-   -   wherein:    -   Y is N or CR¹;    -   Z is N or CR⁴;    -   R¹ is H or trifluoromethyl;    -   R² is selected from the group consisting of H, cyano,        trifluoromethoxy and trifluoromethyl;    -   R³ is selected from the group consisting of H, carboxamide,        chloro, cyano, cyclobutyl, cyclohexyl, cyclopentyl,        cyclopentyloxy, cyclopropyl, cyclopropylmethoxy,        cyclohexylmethyl, 3,3-difluoropyrrolidin-1-yl, ethylamino,        isobutyl, isopropoxy, methylsulfonyl, neopentyl, propyl,        pyrrolidin-1-yl, 1,2,3-thiadiazol-4-yl, trifluoromethoxy and        trifluoromethyl; and    -   R⁴ is selected from the group consisting of H, cyano,        trifluoromethoxy and trifluoromethyl.

Some embodiments of the present invention pertain to compounds selectedfrom compounds of Formula (Ik) and pharmaceutically acceptable salts,solvates and hydrates thereof, wherein:

Y is N or CR¹;

Z is N or CR⁴;

R¹ is H or trifluoromethyl;

R² is selected from the group consisting of H, cyano, trifluoromethoxyand trifluoromethyl;

R³ is selected from the group consisting of H, chloro, cyclobutyl,cyclohexyl, cyclopentyl, cyclopropyl, 3,3-difluoropyrrolidin-1-yl,isobutyl, isopropoxy, neopentyl, propyl, pyrrolidin-1-yl,trifluoromethoxy and trifluoromethyl; and

R⁴ is selected from the group consisting of H, cyano andtrifluoromethyl.

Some embodiments of the present invention pertain to compounds selectedfrom compounds of Formula (Im) and pharmaceutically acceptable salts,solvates and hydrates thereof:

-   -   wherein:    -   R² is selected from the group consisting of H, cyano, C₁-C₆        haloalkoxy and C₁-C₆ haloalkyl; and    -   R³ is selected from the group consisting of H, C₁-C₆ alkoxy,        C₁-C₆ alkyl, C₁-C₆ alkylamino, C₁-C₆ alkylsulfonyl, carboxamide,        cyano, C₃-C₇ cycloalkoxy, C₃-C₇ cycloalkyl, C₁-C₆ haloalkoxy,        C₁-C₆ haloalkyl, halogen, heteroaryl and heterocyclyl, wherein        said C₁-C₆ alkyl and C₁-C₆ alkoxy are each optionally        substituted with one or two substituents selected from C₃-C₇        cycloalkyl and halogen.

Some embodiments of the present invention pertain to compounds selectedfrom compounds of Formula (Im) and pharmaceutically acceptable salts,solvates and hydrates thereof:

wherein:

R² is selected from the group consisting of H, cyano, C₁-C₆ haloalkoxyand C₁-C₆ haloalkyl; and

R³ is selected from the group consisting of H, C₁-C₆ alkoxy, C₁-C₆alkyl, C₃-C₇ cycloalkyl, C₁-C₆ haloalkoxy, C₁-C₆ haloalkyl, halogen andheterocyclyl.

Some embodiments of the present invention pertain to compounds selectedfrom compounds of Formula (Im) and pharmaceutically acceptable salts,solvates and hydrates thereof:

-   -   wherein:    -   R² is selected from the group consisting of H, cyano,        trifluoromethoxy and trifluoromethyl; and    -   R³ is selected from the group consisting of H, carboxamide,        chloro, cyano, cyclobutyl, cyclohexyl, cyclopentyl,        cyclopentyloxy, cyclopropyl, cyclopropylmethoxy,        cyclohexylmethyl, 3,3-difluoropyrrolidin-1-yl, ethylamino,        isobutyl, isopropoxy, methylsulfonyl, neopentyl, propyl,        pyrrolidin-1-yl, 1,2,3-thiadiazol-4-yl, trifluoromethoxy and        trifluoromethyl.

Some embodiments of the present invention pertain to compounds selectedfrom compounds of Formula (Im) and pharmaceutically acceptable salts,solvates and hydrates thereof; wherein:

R² is selected from the group consisting of H, cyano, trifluoromethoxyand trifluoromethyl; and

R³ is selected from the group consisting of H, chloro, cyclobutyl,cyclohexyl, cyclopentyl, cyclopropyl, 3,3-difluoropyrrolidin-1-yl,isobutyl, isopropoxy, neopentyl, propyl, pyrrolidin-1-yl,trifluoromethoxy and trifluoromethyl.

Esters and Prodrugs

One aspect of the present invention pertains to compounds of Formula(IIa) as synthetic intermediates useful in the preparation of compoundsof Formula (Ia) and/or prodrugs useful for the delivery of compounds ofFormula (Ia):

m, n, R², R³, Y, Z and W have the same definitions as described herein,supra and infra, and R⁶ is C₁-C₆ alkyl.

One aspect of the present invention pertains to compounds of Formula(IIa).

In some embodiments, R⁵ is ethyl.

In some embodiments, R⁵ is tert-butyl.

For brevity, it is appreciated that all of the embodiments describedherein, supra and infra, that relate to the common variables sharedbetween Compounds of Formula (Ia) and (IIa) namely, m, n, R², R³, Y, Zand W, apply to Compounds of Formula (IIa) just as if they were eachindividually disclosed herewith with specific reference to Formula(IIa).

One aspect of the present invention pertains to compounds of Formula(IIa) as synthetic intermediates useful in the preparation of compoundsof Formula (Ia).

One aspect of the present invention pertains to compounds of Formula(IIa) as esters of compounds, described and shown herein, such ascompounds in Table A, where R⁵ is ethyl.

One aspect of the present invention pertains to compounds of Formula(IIa) as esters of compounds, described and shown herein, such ascompounds in Table A, where R⁵ is tert-butyl.

One aspect of the present invention pertains to compounds of Formula(IIa) as prodrugs useful for the delivery of compounds of Formula (Ia).

One aspect of the present invention pertains to compounds of Formula(IIa) useful as prodrugs of compounds of Formula (Ia).

Some embodiments of the present invention include every combination ofone or more compounds selected from the following group shown in TableA.

TABLE A Cmpd No. Chemical Structure Chemical Name  1

(S)-2-(7-(3-cyano-4- isopropoxybenzyloxy)-1,2,3,4-tetrahydrocyclopenta[b] indol-3-yl)acetic acid  2

2-(7-(4-cyclohexyl-3- (trifluoromethyl)benzyloxy)- 1,2,3,4-tetrahydrocyclopenta[b] indol-3-yl)acetic acid  3

(R)-2-(7-(4-cyclopentyl-3- (trifluoromethyl)benzyloxy)- 1,2,3,4-tetrahydrocyclopenta[b] indol-3-yl)acetic acid  4

2-(7-(3-cyano-5- (trifluoromethoxy)benzyloxy)- 1,2,3,4-tetrahydrocyclopenta[b] indol-3-yl)acetic acid  5

2-(7-(3-cyano-4- issopropoxybenzyloxy)-1,2,3,4- tetrahydrocyclopenta[b]indol-3-yl)acetic acid  6

(R)-2-(7-(3-cyano-4- isopropoxybenzyloxy)-1,2,3,4-tetrahydrocyclopenta[b] indol-3-yl)acetic acid  7

2-(7-(3-cyano-4- (trifluoromethoxy)benzyloxy)- 1,2,3,4-tetrahydrocyclopenta[b] indol-3-yl)acetic acid  8

2-(7-(2,4- bis(trifluoromethyl)benzyloxy)- 1,2,3,4-tetrahydrocyclopenta[b] indol-3-yl)acetic acid  9

(S)-2-(7-(4-cyclopentyl-3- (trifluoromethyl)benzyloxy)- 1,2,3,4-tetrahydrocyclopenta[b] indol-3-yl)acetic acid 10

2-(7-(3,5- bis(trifluoromethyl)benzyloxy)- 1,2,3,4-tetrahydrocyclopenta[b] indol-3-yl)acetic acid 11

2-(7-((5-isopropoxypyrazin-2- yl)methoxy)-1,2,3,4-tetrahydrocyclopenta[b] indol-3-yl)acetic acid 12

2-(7-(4-cyclopentyl-3- (trifluoromethyl)benzyloxy)- 1,2,3,4-tetrahydrocyclopenta[b] indol-3-yl)acetic acid 13

2-(7-(4-(pyrrolidin-1-yl)-3- (trifluoromethyl)benzyloxy)- 1,2,3,4-tetrahydrocyclopenta[b] indol-3-yl)acetic acid 14

2-(7-(4-isobutyl-3- (trifluoromethyl)benzyloxy)- 1,2,3,4-tetrahydrocyclopenta[b] indol-3-yl)acetic acid 15

2-(7-(4-neopentyl-3- (trifluoromethyl)benzyloxy)- 1,2,3,4-tetrahydrocyclopenta[b] indol-3-yl)acetic acid 16

2-(7-(4-chloro-3- (trifluoromethyl)benzyloxy)- 1,2,3,4-tetrahydrocyclopenta[b] indol-3-yl)acetic acid 17

2-(7-(3-cyano-4- cyclohexylbenzyloxy)-1,2,3,4- tetrahydrocyclopenta[b]indol-3-yl)acetic acid 18

2-(7-(4-propyl-3- (trifluoromethyl)benzyloxy)- 1,2,3,4-tetrahydrocyclopenta[b] indol-3-yl)acetic acid 19

2-(7-((6-cyclopentyl-5- (trifluoromethyl)pyridin-3- yl)methoxy)-1,2,3,4-tetrahydrocyclopenta[b] indol-3-yl)acetic acid 20

2-(7-((6-(3,3- difluoropyrrolidin-1-yl)-5- (trifluoromethyl)pyridin-3-yl)methoxy)-1,2,3,4- tetrahydrocyclopenta[b] indol-3-yl)acetic acid 21

2-(7-(4-cyclobutyl-3- (trifluoromethyl)benzyloxy)- 1,2,3,4-tetrahydrocyclopenta[b] indol-3-yl)acetic acid 22

2-(7-(4-cyclopropyl-3- (trifluoromethyl)benzyloxy)- 1,2,3,4-tetrahydrocyclopenta[b] indol-3-yl)acetic acid 23

2-(7-((6-(pyrrolidin-1-yl)-5- (trifluoromethyl)pyridin-3-yl)methoxy)-1,2,3,4- tetrahydrocyclopenta[b] indol-3-yl)acetic acid 24

2-(7-(4-(cyclopentyloxy)-3- (trifluoromethyl)benzyloxy)- 1,2,3,4-tetrahydrocyclopenta[b]indol-3- yl)acetic acid 25

2-(7-(4-cyano-3- (trifluoromethyl)benzyloxy)- 1,2,3,4-tetrahydrocyclopenta[b]indol-3- yl)acetic acid 26

2-(7-(4-(cyclopropylmethyl)- 3-(trifluoromethyl)benzyloxy)- 1,2,3,4-tetrahydrocyclopenta[b]indol-3- yl)acetic acid 27

2-(7-(4-(ethylamino)-3- (trifluoromethyl)benzyloxy)- 1,2,3,4-tetrahydrocyclopenta[b]indol-3- yl)acetic acid 28

2-(7-(4-(cyclohexylmethyl)-3- (trifluoromethyl)benzyloxy)- 1,2,3,4-tetrahydrocyclopenta[b]indol-3- yl)acetic acid 29

2-(7-(4-carbamoyl-3- (trifluoromethyl)benzyloxy)- 1,2,3,4-tetrahydrocyclopenta[b]indol-3- yl)acetic acid 30

2-(7-(4- (methylsulfonyl)benzyloxy)- 1,2,3,4-tetrahydrocyclopenta[b]indol-3- yl)acetic acid 31

2-(7-(4-(pyrazin-2-yl)- benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3- yl)acetic acid 32

2-(7-(4-(1,2,3-thiadiazol-4- yl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3- yl)acetic acid

Additionally, individual compounds and chemical genera of the presentinvention, for example, those compounds found in Table A includingdiastereomers and enantiomers thereof, encompass all pharmaceuticallyacceptable salts, solvates and hydrates, thereof.

It is understood that the present invention embraces each diastereomer,each enantiomer and mixtures thereof of each compound and genericformulae disclosed herein just as if they were each individuallydisclosed with the specific stereochemical designation for each chiralcarbon. Separation of the individual isomers (such as, by chiral HPLC,recrystallization of diastereomeric mixtures and the like) or selectivesynthesis (such as, by enantiomeric selective syntheses and the like) ofthe individual isomers is accomplished by application of various methodswhich are well known to practitioners in the art.

The compounds of the Formula (Ia) of the present invention may beprepared according to relevant published literature procedures that areused by one skilled in the art. Exemplary reagents and procedures forthese reactions appear hereinafter in the working examples. Protectionand deprotection may be carried out by procedures generally known in theart (see, for example, Greene, T. W. and Wuts, P. G. M., ProtectingGroups in Organic Synthesis, 3^(rd) Edition, 1999 [Wiley]; incorporatedherein by reference in its entirety).

The embodiments of the present invention include every combination ofone or more salts selected from the following group and pharmaceuticallyacceptable solvates and hydrates thereof:

-   Calcium salt of    (R)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetic    acid; and-   L-Arginine salt of    (R)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetic    acid.

Pharmaceutical Compositions

A further aspect of the present invention pertains to pharmaceuticalcompositions comprising one or more compounds as described herein andone or more pharmaceutically acceptable carriers. Some embodimentspertain to pharmaceutical compositions comprising a compound of thepresent invention and a pharmaceutically acceptable carrier.

Some embodiments of the present invention include a method of producinga pharmaceutical composition comprising admixing at least one compoundaccording to any of the compound embodiments disclosed herein and apharmaceutically acceptable carrier.

Formulations may be prepared by any suitable method, typically byuniformly mixing the active compound(s) with liquids or finely dividedsolid carriers, or both, in the required proportions and then, ifnecessary, forming the resulting mixture into a desired shape.

Conventional excipients, such as binding agents, fillers, acceptablewetting agents, tabletting lubricants and disintegrants may be used intablets and capsules for oral administration. Liquid preparations fororal administration may be in the form of solutions, emulsions, aqueousor oily suspensions and syrups. Alternatively, the oral preparations maybe in the form of dry powder that can be reconstituted with water oranother suitable liquid vehicle before use. Additional additives such assuspending or emulsifying agents, non-aqueous vehicles (including edibleoils), preservatives and flavorings and colorants may be added to theliquid preparations. Parenteral dosage forms may be prepared bydissolving the compound of the invention in a suitable liquid vehicleand filter sterilizing the solution before filling and sealing anappropriate vial or ampoule. These are just a few examples of the manyappropriate methods well known in the art for preparing dosage forms.

A compound of the present invention can be formulated intopharmaceutical compositions using techniques well known to those in theart. Suitable pharmaceutically acceptable carriers, outside thosementioned herein, are known in the art; for example, see Remington, TheScience and Practice of Pharmacy, 20^(th) Edition, 2000, LippincottWilliams & Wilkins, (Editors: Gennaro et al.)

While it is possible that, for use in the prophylaxis or treatment, acompound of the invention may, in an alternative use, be administered asa raw or pure chemical, it is preferable however to present the compoundor active ingredient as a pharmaceutical formulation or compositionfurther comprising a pharmaceutically acceptable carrier.

The invention thus further provides pharmaceutical formulationscomprising a compound of the invention or a pharmaceutically acceptablesalt, solvate, hydrate or derivative thereof together with one or morepharmaceutically acceptable carriers thereof and/or prophylacticingredients. The carrier(s) must be “acceptable” in the sense of beingcompatible with the other ingredients of the formulation and not overlydeleterious to the recipient thereof.

Pharmaceutical formulations include those suitable for oral, rectal,nasal, topical (including buccal and sub-lingual), vaginal or parenteral(including intramuscular, subcutaneous and intravenous) administrationor in a form suitable for administration by inhalation, insufflation orby a transdermal patch. Transdermal patches dispense a drug at acontrolled rate by presenting the drug for absorption in an efficientmanner with a minimum of degradation of the drug. Typically, transdermalpatches comprise an impermeable backing layer, a single pressuresensitive adhesive and a removable protective layer with a releaseliner. One of ordinary skill in the art will understand and appreciatethe techniques appropriate for manufacturing a desired efficacioustransdermal patch based upon the needs of the artisan.

The compounds of the invention, together with a conventional adjuvant,carrier, or diluent, may thus be placed into the form of pharmaceuticalformulations and unit dosages thereof and in such form may be employedas solids, such as tablets or filled capsules, or liquids such assolutions, suspensions, emulsions, elixirs, gels or capsules filled withthe same, all for oral use; in the form of suppositories for rectaladministration; or in the form of sterile injectable solutions forparenteral (including subcutaneous) use. Such pharmaceuticalcompositions and unit dosage forms thereof may comprise conventionalingredients in conventional proportions, with or without additionalactive compounds or principles and such unit dosage forms may containany suitable effective amount of the active ingredient commensurate withthe intended daily dosage range to be employed.

For oral administration, the pharmaceutical composition may be in theform of, for example, a tablet, capsule, suspension or liquid. Thepharmaceutical composition is preferably made in the form of a dosageunit containing a particular amount of the active ingredient. Examplesof such dosage units are capsules, tablets, powders, granules orsuspensions, with conventional additives such as lactose, mannitol, cornstarch or potato starch; with binders such as crystalline cellulose,cellulose derivatives, acacia, corn starch or gelatins; withdisintegrators such as corn starch, potato starch or sodiumcarboxymethyl-cellulose; and with lubricants such as talc or magnesiumstearate. The active ingredient may also be administered by injection asa composition wherein, for example, saline, dextrose or water may beused as a suitable pharmaceutically acceptable carrier.

Compounds of the present invention or a salt, solvate, hydrate orphysiologically functional derivative thereof can be used as activeingredients in pharmaceutical compositions, specifically as S1P1receptor modulators. The term “active ingredient” is defined in thecontext of a “pharmaceutical composition” and is intended to mean acomponent of a pharmaceutical composition that provides the primarypharmacological effect, as opposed to an “inactive ingredient” whichwould generally be recognized as providing no pharmaceutical benefit.

The dose when using the compounds of the present invention can varywithin wide limits and as is customary and known to the physician, it isto be tailored to the individual conditions in each individual case. Itdepends, for example, on the nature and severity of the illness to betreated, on the condition of the patient, on the compound employed or onwhether an acute or chronic disease state is treated or prophylaxis isconducted or on whether further active compounds are administered inaddition to the compounds of the present invention. Representative dosesof the present invention include, but are not limited to, about 0.001 mgto about 5000 mg, about 0.001 mg to about 2500 mg, about 0.001 mg toabout 1000 mg, 0.001 mg to about 500 mg, 0.001 mg to about 250 mg, about0.001 mg to 100 mg, about 0.001 mg to about 50 mg and about 0.001 mg toabout 25 mg. Multiple doses may be administered during the day,especially when relatively large amounts are deemed to be needed, forexample 2, 3 or 4 doses. Depending on the individual and as deemedappropriate by the patient's physician or caregiver it may be necessaryto deviate upward or downward from the doses described herein.

The amount of active ingredient or an active salt, solvate or hydratederivative thereof; required for use in treatment will vary not onlywith the particular salt selected but also with the route ofadministration, the nature of the condition being treated and the ageand condition of the patient and will ultimately be at the discretion ofthe attendant physician or clinician. In general, one skilled in the artunderstands how to extrapolate in vivo data obtained in one modelsystem, typically an animal model, to another, such as a human. In somecircumstances, these extrapolations may merely be based on the weight ofthe animal model in comparison to another, such as a mammal, preferablya human, however, more often, these extrapolations are not simply basedon weights, but rather incorporate a variety of factors. Representativefactors include the type, age, weight, sex, diet and medical conditionof the patient, the severity of the disease, the route ofadministration, pharmacological considerations such as the activity,efficacy, pharmacokinetic and toxicology profiles of the particularcompound employed, whether a drug delivery system is utilized, whetheran acute or chronic disease state is being treated or prophylaxis isconducted or whether further active compounds are administered inaddition to the compounds of the present invention and as part of a drugcombination. The dosage regimen for treating a disease condition withthe compounds and/or compositions of this invention is selected inaccordance with a variety factors including those cited above. Thus, theactual dosage regimen employed may vary widely and therefore may deviatefrom a preferred dosage regimen and one skilled in the art willrecognize that dosage and dosage regimens outside these typical rangescan be tested and, where appropriate, may be used in the methods of thisinvention.

The desired dose may conveniently be presented in a single dose or asdivided doses administered at appropriate intervals, for example, as 2,3, 4 or more sub-doses per day. The sub-dose itself may be furtherdivided, e.g., into a number of discrete loosely spaced administrations.The daily dose can be divided, especially when relatively large amountsare administered as deemed appropriate, into several, for example 2, 3or 4 part administrations. If appropriate, depending on individualbehavior, it may be necessary to deviate upward or downward from thedaily dose indicated.

For preparing pharmaceutical compositions from the compounds of thepresent invention, the suitable pharmaceutically acceptable carrier canbe either solid, liquid or a mixture of both. Solid form preparationsinclude powders, tablets, pills, capsules, cachets, suppositories anddispersible granules. A solid carrier can be one or more substanceswhich may also act as diluents, flavoring agents, solubilizers,lubricants, suspending agents, binders, preservatives, tabletdisintegrating agents, or encapsulating materials.

In powders, the carrier is a finely divided solid which is in a mixturewith the finely divided active component.

In tablets, the active component is mixed with the carrier having thenecessary binding capacity in suitable proportions and compacted to thedesired shape and size.

The powders and tablets may contain varying percentage amounts of theactive compound. A representative amount in a powder or tablet may befrom 0.5 to about 90 percent of the active compound. However, an artisanwould know when amounts outside of this range are necessary. Suitablecarriers for powders and tablets include magnesium carbonate, magnesiumstearate, talc, sugar, lactose, pectin, dextrin, starch, gelatin,tragacanth, methylcellulose, sodium carboxymethylcellulose, a lowmelting wax, cocoa butter and the like. The term “preparation” isintended to include the formulation of the active compound withencapsulating material as carrier providing a capsule in which theactive component, with or without carriers, is surrounded by a carrier,which is thus in association with it. Similarly, cachets and lozengesare included. Tablets, powders, capsules, pills, cachets and lozengescan be used as solid forms suitable for oral administration.

For preparing suppositories, a low melting wax, such as an admixture offatty acid glycerides or cocoa butter, is first melted and the activecomponent is dispersed homogeneously therein (e.g., by stirring). Themolten homogenous mixture is then poured into convenient sized molds,allowed to cool and thereby to solidify.

Formulations suitable for vaginal administration may be presented aspessaries, tampons, creams, gels, pastes, foams or sprays containing inaddition to the active ingredient such carriers as are known in the artto be appropriate.

Liquid form preparations include solutions, suspensions and emulsions,for example, water or water-propylene glycol solutions. For example,parenteral injection liquid preparations can be formulated as solutionsin aqueous polyethylene glycol solution. Injectable preparations, forexample, sterile injectable aqueous or oleaginous suspensions may beformulated according to the known art using suitable dispersing orwetting agents and suspending agents. The sterile injectable preparationmay also be a sterile injectable solution or suspension in a nontoxicparenterally acceptable diluent or solvent, for example, as a solutionin 1,3-butanediol. Among the acceptable vehicles and solvents that maybe employed are water, Ringer's solution and isotonic sodium chloridesolution. In addition, sterile, fixed oils are conventionally employedas a solvent or suspending medium. For this purpose any bland fixed oilmay be employed including synthetic mono- or diglycerides. In addition,fatty acids such as oleic acid find use in the preparation ofinjectables.

The compounds according to the present invention may thus be formulatedfor parenteral administration (e.g. by injection, for example bolusinjection or continuous infusion) and may be presented in unit dose formin ampoules, pre-filled syringes, small volume infusion or in multi-dosecontainers with an added preservative. The pharmaceutical compositionsmay take such forms as suspensions, solutions, or emulsions in oily oraqueous vehicles and may contain formulatory agents such as suspending,stabilizing and/or dispersing agents. Alternatively, the activeingredient may be in powder form, obtained by aseptic isolation ofsterile solid or by lyophilization from solution, for constitution witha suitable vehicle, e.g. sterile, pyrogen-free water, before use.

Aqueous formulations suitable for oral use can be prepared by dissolvingor suspending the active component in water and adding suitablecolorants, flavors, stabilizing and thickening agents, as desired.

Aqueous suspensions suitable for oral use can be made by dispersing thefinely divided active component in water with viscous material, such asnatural or synthetic gums, resins, methylcellulose, sodiumcarboxymethylcellulose, or other well-known suspending agents.

Also included are solid form preparations which are intended to beconverted, shortly before use, to liquid form preparations for oraladministration. Such liquid forms include solutions, suspensions andemulsions. These preparations may contain, in addition to the activecomponent, colorants, flavors, stabilizers, buffers, artificial andnatural sweeteners, dispersants, thickeners, solubilizing agents and thelike.

For topical administration to the epidermis the compounds according tothe invention may be formulated as ointments, creams or lotions, or as atransdermal patch.

Ointments and creams may, for example, be formulated with an aqueous oroily base with the addition of suitable thickening and/or gellingagents. Lotions may be formulated with an aqueous or oily base and willin general also contain one or more emulsifying agents, stabilizingagents, dispersing agents, suspending agents, thickening agents, orcoloring agents.

Formulations suitable for topical administration in the mouth includelozenges comprising the active agent in a flavored base, usually sucroseand acacia or tragacanth; pastilles comprising the active ingredient inan inert base such as gelatin and glycerin or sucrose and acacia; andmouthwashes comprising the active ingredient in a suitable liquidcarrier.

Solutions or suspensions are applied directly to the nasal cavity byconventional means, for example with a dropper, pipette or spray. Theformulations may be provided in single or multi-dose form. In the lattercase of a dropper or pipette, this may be achieved by the patientadministering an appropriate, predetermined volume of the solution orsuspension. In the case of a spray, this may be achieved for example bymeans of a metering atomizing spray pump.

Administration to the respiratory tract may also be achieved by means ofan aerosol formulation in which the active ingredient is provided in apressurized pack with a suitable propellant. If the compounds of thepresent invention or pharmaceutical compositions comprising them areadministered as aerosols (e.g., nasal aerosols, by inhalation), this canbe carried out, for example, using a spray, a nebulizer, a pumpnebulizer, an inhalation apparatus, a metered inhaler or a dry powderinhaler. Pharmaceutical forms for administration of the compounds of thepresent invention as an aerosol can be prepared by processes well knownto the person skilled in the art. Solutions or dispersions of thecompounds of the present invention or a pharmaceutically acceptablesalt, solvate, hydrate or derivative thereof in water, water/alcoholmixtures or suitable saline solutions, for example, can be employedusing customary additives (e.g., benzyl alcohol or other suitablepreservatives), absorption enhancers for increasing the bioavailability,solubilizers, dispersants and others and, if appropriate, customarypropellants (e.g., carbon dioxide, CFCs, such as,dichlorodifluoromethane, trichlorofluoromethane,dichlorotetrafluoroethane and the like). The aerosol may convenientlyalso contain a surfactant such as lecithin. The dose of drug may becontrolled by provision of a metered valve.

In formulations intended for administration to the respiratory tract,including intranasal formulations, the compound will generally have asmall particle size for example of the order of 10 microns or less. Sucha particle size may be obtained by means known in the art, for exampleby micronization. When desired, formulations adapted to give sustainedrelease of the active ingredient may be employed.

Alternatively the active ingredients may be provided in the form of adry powder (e.g., a powder mix of the compound in a suitable powder basesuch as lactose, starch, starch derivatives such as hydroxypropylmethylcellulose and polyvinylpyrrolidone (PVP)). Conveniently the powdercarrier will form a gel in the nasal cavity. The powder composition maybe presented in unit dose form (e.g., capsules, cartridges) as forgelatin or blister packs from which the powder may be administered bymeans of an inhaler.

The pharmaceutical preparations are preferably in unit dosage forms. Insuch form, the preparation is subdivided into unit doses containingappropriate quantities of the active component. The unit dosage form canbe a packaged preparation, the package containing discrete quantities ofpreparation, such as packeted tablets, capsules and powders in vials orampoules. Also, the unit dosage form can be a capsule, tablet, cachet,or lozenge itself, or it can be the appropriate number of any of thesein packaged form.

In some embodiments, the compositions are tablets or capsules for oraladministration.

In some embodiments, the compositions are liquids for intravenousadministration.

The compounds according to the invention may optionally exist aspharmaceutically acceptable salts including pharmaceutically acceptableacid addition salts prepared from pharmaceutically acceptable non-toxicacids including inorganic and organic acids. Representative acidsinclude, but are not limited to, acetic, benzenesulfonic, benzoic,camphorsulfonic, citric, ethenesulfonic, dichloroacetic, formic,fumaric, gluconic, glutamic, hippuric, hydrobromic, hydrochloric,isethionic, lactic, maleic, malic, mandelic, methanesulfonic, mucic,nitric, oxalic, pamoic, pantothenic, phosphoric, succinic, sulfuric,tartaric, oxalic, p-toluenesulfonic and the like, such as thosepharmaceutically acceptable salts listed by Berge et al., Journal ofPharmaceutical Sciences, 66:1-19 (1977), incorporated herein byreference in its entirety.

The acid addition salts may be obtained as the direct products ofcompound synthesis. In the alternative, the free base may be dissolvedin a suitable solvent containing the appropriate acid and the saltisolated by evaporating the solvent or otherwise separating the salt andsolvent. The compounds of this invention may form solvates with standardlow molecular weight solvents using methods known to the skilledartisan.

Compounds of the present invention can be converted to “pro-drugs.” Theterm “pro-drugs” refers to compounds that have been modified withspecific chemical groups known in the art and that when administeredinto an individual undergo biotransformation to give the parentcompound. Pro-drugs can thus be viewed as compounds of the inventioncontaining one or more specialized non-toxic protective groups used in atransient manner to alter or to eliminate a property of the compound. Inone general aspect, the “pro-drug” approach is utilized to facilitateoral absorption. A thorough discussion is provided in T. Higuchi and V.Stella, Pro-drugs as Novel Delivery Systems Vol. 14 of the A.C.S.Symposium Series; and in Bioreversible Carriers in Drug Design, ed.Edward B. Roche, American Pharmaceutical Association and Pergamon Press,1987, both of which are hereby incorporated by reference in theirentirety.

Some embodiments of the present invention include a method of producinga pharmaceutical composition for “combination-therapy” comprisingadmixing at least one compound according to any of the compoundembodiments disclosed herein, together with at least one knownpharmaceutical agent as described herein and a pharmaceuticallyacceptable carrier.

It is noted that when S1P1 receptor agonists are utilized as activeingredients in a pharmaceutical composition, these are not intended foruse only in humans, but in other non-human mammals as well. Indeed,recent advances in the area of animal health-care mandate thatconsideration be given for the use of active agents, such as S1P1receptor agonists, for the treatment of an S1P1 receptor-associateddisease or disorder in companionship animals (e.g., cats, dogs, etc.)and in livestock animals (e.g., cows, chickens, fish, etc.). Those ofordinary skill in the art are readily credited with understanding theutility of such compounds in such settings.

Hydrates and Solvates

It is understood that when the phrase “pharmaceutically acceptablesalts, solvates and hydrates” is used in reference to a particularformula herein, it is intended to embrace solvates and/or hydrates ofcompounds of the particular formula, pharmaceutically acceptable saltsof compounds of the particular formula as well as solvates and/orhydrates of pharmaceutically acceptable salts of compounds of theparticular formula. It is also understood by a person of ordinary skillin the art that hydrates are a subgenus of solvates.

The compounds of the present invention can be administrated in a widevariety of oral and parenteral dosage forms. It will be apparent tothose skilled in the art that the following dosage forms may comprise,as the active component, either a compound of the invention or apharmaceutically acceptable salt or as a solvate or hydrate thereof.Moreover, various hydrates and solvates of the compounds of theinvention and their salts will find use as intermediates in themanufacture of pharmaceutical compositions. Typical procedures formaking and identifying suitable hydrates and solvates, outside thosementioned herein, are well known to those in the art; see for example,pages 202-209 of K. J. Guillory, “Generation of Polymorphs, Hydrates,Solvates, and Amorphous Solids,” in: Polymorphism in PharmaceuticalSolids, ed. Harry G. Brittan, Vol. 95, Marcel Dekker, Inc., New York,1999, incorporated herein by reference in its entirety. Accordingly, oneaspect of the present invention pertains to hydrates and solvates ofcompounds of the present invention and/or their pharmaceuticalacceptable salts, as described herein, that can be isolated andcharacterized by methods known in the art, such as, thermogravimetricanalysis (TGA), TGA-mass spectroscopy, TGA-Infrared spectroscopy, powderX-ray diffraction (PXRD), Karl Fisher titration, high resolution X-raydiffraction, and the like. There are several commercial entities thatprovide quick and efficient services for identifying solvates andhydrates on a routine basis. Example companies offering these servicesinclude Wilmington PharmaTech (Wilmington, Del.), Avantium Technologies(Amsterdam) and Aptuit (Greenwich, Conn.).

The embodiments of the present invention include every combination ofone or more solvate or hydrate selected from the following group:

-   D-Lysine salt of    (S)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetic    acid hydrate; and-   (R)-1-Phenethylamine salt of    (S)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetic    acid acetonitrile solvate.

In some embodiments, the crystalline form is(S)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)aceticacid hydrate.

In some embodiments, the crystalline form of(S)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)aceticacid hydrate has an X-ray powder diffraction pattern substantially asshown in FIG. 12, wherein by “substantially” is meant that the reportedpeaks can vary by about ±0.2 °2θ.

In some embodiments, the crystalline form of(S)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)aceticacid hydrate has a differential scanning calorimetry thermogramsubstantially as shown in FIG. 13, wherein by “substantially” is meantthat the reported DSC features can vary by about ±4° C.

In some embodiments, the crystalline form of(S)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)aceticacid hydrate has a thermogravimetric analysis thermogram substantiallyas shown in FIG. 13.

In some embodiments, the crystalline form of(S)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)aceticacid hydrate has a moisture-sorption analysis substantially as shown inFIG. 14, wherein by “substantially” is meant that the reportedmoisture-sorption analysis features can vary by about ±5% relativehumidity.

Other Utilities

Another object of the present invention relates to radiolabeledcompounds of the present invention that are useful not only inradio-imaging but also in assays, both in vitro and in vivo, forlocalizing and quantitating the S1P1 receptor in tissue samples,including human and for identifying S1P1 receptor ligands by inhibitionbinding of a radiolabeled compound. It is a further object of thisinvention to develop novel S1P1 receptor assays which comprise suchradiolabeled compounds.

The present invention embraces isotopically-labeled compounds of thepresent invention. Isotopically or radiolabeled compounds are thosewhich are identical to compounds disclosed herein, but for the fact thatone or more atoms are replaced or substituted by an atom having anatomic mass or mass number different from the atomic mass or mass numbermost commonly found in nature. Suitable radionuclides that may beincorporated in compounds of the present invention include, but are notlimited, to ²H (also written as D for deuterium), ³H (also written as Tfor tritium), ¹¹C, ¹³C, ¹⁴C, ¹³N, ¹⁵N, ¹⁵O, ¹⁷O, ¹⁸O, ¹⁸F, ³⁵S, ³⁶Cl,⁷⁵Br, ⁷⁶Br, ⁷⁷Br, ⁸²Br, ¹²³I, ¹²⁴I, ¹²⁵I and ¹³¹I. The radionuclide thatis incorporated in the instant radiolabeled compounds will depend on thespecific application of that radiolabeled compound. For example, for invitro S1P1 receptor labeling and competition assays, compounds thatincorporate ³H, ¹⁴C, ⁸²Br, ¹²⁵I, ¹³¹I or ³⁵S will generally be mostuseful. For radio-imaging applications ¹¹C, ¹⁸F, ¹²⁵I, ¹²³I, ¹²⁴I, ¹³¹I,⁷⁵Br, ⁷⁶Br or ⁷⁷Br will generally be most useful.

It is understood that a “radiolabeled” or “labeled compound” is acompound of Formula (Ia), (Ic), (Ie), (Ig), (Ii), (Ik) or (Im)containing at least one radionuclide. In some embodiments theradionuclide is selected from the group consisting of ³H, ¹⁴C, ¹²⁵I, ³⁵Sand ⁸²Br.

Certain isotopically-labeled compounds of the present invention areuseful in compound and/or substrate tissue distribution assays. In someembodiments the radionuclide ³H and/or ¹⁴C isotopes are useful in thesestudies. Further, substitution with heavier isotopes such as deuterium(i.e., ²H) may afford certain therapeutic advantages resulting fromgreater metabolic stability (e.g., increased in vivo half-life orreduced dosage requirements) and hence may be preferred in somecircumstances. Isotopically labeled compounds of the present inventioncan generally be prepared by following procedures analogous to thosedisclosed in FIGS. 3 to 6 and examples infra, by substituting anisotopically labeled reagent for a non-isotopically labeled reagent.Other synthetic methods that are useful are discussed infra. Moreover,it should be understood that all of the atoms represented in thecompounds of the invention can be either the most commonly occurringisotope of such atoms or a scarcer radio-isotope or nonradioactiveisotope.

Synthetic methods for incorporating radio-isotopes into organiccompounds are applicable to compounds of the invention and are wellknown in the art. Certain synthetic methods, for example, forincorporating activity levels of tritium into target molecules, are asfollows:

A. Catalytic Reduction with Tritium Gas: This procedure normally yieldshigh specific activity products and requires halogenated or unsaturatedprecursors.

B. Reduction with Sodium Borohydride [³H]: This procedure is ratherinexpensive and requires precursors containing reducible functionalgroups such as aldehydes, ketones, lactones, esters and the like.

C. Reduction with Lithium Aluminum Hydride [³H]: This procedure offersproducts at almost theoretical specific activities. It also requiresprecursors containing reducible functional groups such as aldehydes,ketones, lactones, esters and the like.

D. Tritium Gas Exposure Labeling: This procedure involves exposingprecursors containing exchangeable protons to tritium gas in thepresence of a suitable catalyst.

E. N-Methylation using Methyl Iodide [³H]: This procedure is usuallyemployed to prepare O-methyl or N-methyl [³H] products by treatingappropriate precursors with high specific activity methyl iodide [³H].This method in general allows for higher specific activity, such as forexample, about 70-90 Ci/mmol.

Synthetic methods for incorporating activity levels of ¹²⁵I into targetmolecules include:

A. Sandmeyer and like reactions: This procedure transforms an aryl amineor a heteroaryl amine into a diazonium salt, such as a diazoniumtetrafluoroborate salt and subsequently to ¹²⁵I labeled compound usingNa¹²⁵I. A represented procedure was reported by Zhu, G-D. and co-workersin J. Org. Chem., 2002, 67, 943-948.

B. Ortho ¹²⁵Iodination of phenols: This procedure allows for theincorporation of ¹²⁵I at the ortho position of a phenol as reported byCollier, T. L. and co-workers in J. Labelled Compd. Radiopharm., 1999,42, S264-S266.

C. Aryl and heteroaryl bromide exchange with ¹²⁵I: This method isgenerally a two step process. The first step is the conversion of thearyl or heteroaryl bromide to the corresponding tri-alkyltinintermediate using for example, a Pd catalyzed reaction [i.e. Pd(Ph₃P)₄]or through an aryl or heteroaryl lithium, in the presence of atri-alkyltinhalide or hexaalkylditin [e.g., (CH₃)₃SnSn(CH₃)₃]. Arepresentative procedure was reported by Le Bas, M.-D. and co-workers inJ. Labelled Compd. Radiopharm. 2001, 44, S280-S282.

A radiolabeled S1P1 receptor compound of Formula (Ia) can be used in ascreening assay to identify/evaluate compounds. In general terms, anewly synthesized or identified compound (i.e., test compound) can beevaluated for its ability to reduce binding of the “radiolabeledcompound of Formula (Ia)” to the S1P1 receptor. Accordingly, the abilityof a test compound to compete with the “radiolabeled compound of Formula(Ia)” for the binding to the S1P1 receptor directly correlates to itsbinding affinity.

The labeled compounds of the present invention bind to the S1P1receptor. In one embodiment the labeled compound has an IC₅₀ less thanabout 500 μM, in another embodiment the labeled compound has an IC₅₀less than about 100 μM, in yet another embodiment the labeled compoundhas an IC₅₀ less than about 10 μM, in yet another embodiment the labeledcompound has an IC₅₀ less than about 1 μM and in still yet anotherembodiment the labeled inhibitor has an IC₅₀ less than about 0.1 μM.

Other uses of the disclosed receptors and methods will become apparentto those of skill in the art based upon, inter alia, a review of thisdisclosure.

As will be recognized, the steps of the methods of the present inventionneed not be performed any particular number of times or in anyparticular sequence. Additional objects, advantages and novel featuresof this invention will become apparent to those skilled in the art uponexamination of the following examples thereof, which are intended to beillustrative and not intended to be limiting.

EXAMPLES Example 1 Syntheses of Compounds of the Present Invention

Illustrated syntheses for compounds of the present invention are shownin FIGS. 3 through 6 where the variables have the same definitions asused throughout this disclosure.

The compounds of the invention and their syntheses are furtherillustrated by the following examples. The following examples areprovided to further define the invention without, however, limiting theinvention to the particulars of these examples. The compounds describedherein, supra and infra, are named according to the AutoNom version 2.2,CS ChemDraw Ultra Version 9.0.7. In certain instances common names areused and it is understood that these common names would be recognized bythose skilled in the art.

Chemistry:

Proton nuclear magnetic resonance (¹H NMR) spectra were recorded on aBruker Avance-400 equipped with a QNP (Quad Nucleus Probe) or a BBI(Broad Band Inverse) and z-gradient. Proton nuclear magnetic resonance(¹H NMR) spectra were also recorded on a Broker Avance-500 equipped aBBI (Broad Band Inverse) and z-gradient. Chemical shifts are given inparts per million (ppm) with the residual solvent signal used asreference. NMR abbreviations are used as follows: s=singlet, d=doublet,dd=doublet of doublets, t=triplet, q=quartet, m=multiplet, bs=broadsinglet. Microwave irradiations were carried out using a SmithSynthesizer™ or an Emrys Optimizer™ (Biotage). Thin-layer chromatography(TLC) was performed on silica gel 60 F₂₅₄ (Merck), preparatorythin-layer chromatography (prep TLC) was preformed on PK6F silica gel 60A 1 mm plates (Whatman) and column chromatography was carried out on asilica gel column using Kieselgel 60, 0.063-0.200 mm (Merck).Evaporation was done under reduced pressure on a Büchi rotaryevaporator. Celite® 545 was used for filtration of palladium.

LCMS spec: HPLC-pumps: LC-LOAD VP, Shimadzu Inc.; HPLC systemcontroller: SCL-10A VP, Shimadzu Inc; UV-Detector: SPD-10A VP, ShimadzuInc; Autosampler: CTC HTS, PAL, Leap Scientific; Mass spectrometer: API150EX with Turbo Ion Spray source, AB/MDS Sciex; Software: Analyst 1.2.

Example 1.1 Preparation of2-(7-(3-Cyano-5-(trifluoromethoxy)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)aceticAcid (Compound 4) Step A: Preparation of Ethyl1-(2-Ethoxy-2-oxoethyl)-2-oxocyclopentanecarboxylate

To a solution of ethyl 2-oxocyclopentanecarboxylate (93.27 g, 597 mmol)and ethyl 2-bromoacetate (144.64 g, 866 mmol) in acetone (1.2 L) wasadded K₂CO₃ (165 g, 1194 mmol). The mixture was heated at 56° C. for 24h. The solid was filtered off and the filtering cake was washed withacetone (3×100 mL). The filtrate was concentrated and the resultantliquid was purified by a silica gel plug to give the title compound aslight yellow liquid (54.7 g). LCMS m/z=243.3 [M+H]⁺; ¹H NMR (400 MHz,CDCl₃) δ ppm 1.23 (t, J=7.14 Hz, 3H), 1.24 (t, J=7.14 Hz, 3H), 1.95-2.03(m, 1H), 2.06-2.15 (m, 2H), 2.35-2.50 (m, 2H), 2.55-2.60 (m, 1H), 2.80(dd, J=15.2, 2.09 Hz, 1H), 2.95 (dd, J=15.2, 2.09 Hz, 1H), 4.09 (q,J=7.14 Hz, 2H), 4.12 (q, J=7.14 Hz, 2H).

Step B: Preparation of 2-(2-Oxocyclopentyl)acetic Acid

A solution of ethyl 1-(2-ethoxy-2-oxoethyl)-2-oxocyclopentanecarboxylate(50.0 g, 206 mmol) in HOAc (500 mL) and 6 M HCl (250 mL) was heated at100° C. for 6 h. The solvent was removed under reduced pressure and theresidue was partitioned between EtOAc (500 mL) and H₂O (200 mL). Aqueouslayer was separated and extracted with EtOAc (2×250 mL). The combinedorganic layers were washed with H₂O (300 mL), brine (300 mL), dried overNa₂SO₄, decanted and concentrated to yield the title compound as a whitesolid (22 g). ¹H NMR (400 MHz, CDCl₃) δ ppm 1.59-1.72 (m, 1H), 1.75-1.90(m, 1H), 2.03-2.10 (m, 1H), 2.20 (dd, J=10.9, 8.9 Hz, 1H), 2.30-2.40 (m,2H), 2.40-2.50 (m, 2H), 2.80 (dd, J=15.7, 7.2 Hz, 1H), 11.5 (s, 1H).

Step C: Preparation of Ethyl 2-(2-Oxocyclopentyl)acetate

To a solution of 2-(2-oxocyclopentyl)acetic acid (23.6 g, 166 mmol) inabsolute ethanol (400 mL) was added H₂SO₄ (16.28 g, 166 mmol). Theresultant solution was heated under reflux overnight. The reactionmixture was concentrated and the liquid residue was added into ice-water(200 mL). The aqueous mixture was extracted with DCM (3×200 mL). Thecombined organic layers were washed with H₂O (300 mL), brine (300 mL),dried over Na₂SO₄, decanted, concentrated and dried under vacuum toafford the title compound as a light yellow liquid (27.2 g). LCMSm/z=171.3 [M+H]⁺; ¹H NMR (400 MHz, CDCl₃) δ ppm 1.19 (t, J=7.14 Hz, 3H),1.50-1.62 (m, 1H), 1.65-1.80 (m, 1H), 1.92-2.02 (m, 1H), 2.12 (dd,J=16.7, 8.86 Hz, 1H), 2.19-2.29 (m, 2H), 2.30-2.44 (m, 2H), 2.65 (dd,J=15.12, 2.6 Hz, 1H), 4.07 (q, J=7.14 Hz, 2H).

Step D: Preparation of Ethyl2-(7-Hydroxy-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetate

2-Iodo-4-methoxyaniline (2.0 g, 8.03 mmol) and ethyl2-(2-oxocyclopentyl)acetate (2.05 g, 12.1 mmol) were dissolved in DMF(30 mL) and tetraethyl orthosilicate (2.12 g, 10.4 mmol) and pyridiniump-toluenesulfonate (PPTS) (0.081 g, 0.321 mmol) were added. The reactionmixture was heated and stirred at 135° C. for 4 h. After cooling to 120°C., DIEA (3.11 g, 24.09 mmol) and palladium (II) acetate (0.054 g, 0.241mmol) were added. The reaction mixture was stirred for 3 h and thenpartitioned between ethyl acetate and water. The aqueous layer wasextracted twice with ethyl acetate. The combined organic extracts weredried over sodium sulfate, filtered and concentrated under reducedpressure. The resultant solution was diluted with 50% ethyl acetate inhexanes and filtered through a pad of silica gel. The filtrate wasconcentrated and purified by silica gel column chromatography to give1.9 g of ethyl2-(7-methoxy-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetatecontaining residual ethyl 2-(2-oxocyclopentyl)acetate. The mixture wasdissolved in DCM (80 mL) and cooled to 0° C. Boron tribromide (21.0 mL,21.0 mmol, 1.0 M in DCM) was added and the reaction was stirred for 1.5h. Ice water was added and the reaction mixture was allowed to reachroom temperature. The aqueous mixture was extracted three times withDCM. The combined organics were dried over sodium sulfate, filtered andconcentrated under reduced pressure. The residue was purified by silicagel column chromatography to give the title compound (650 mg). LCMSm/z=260.3 [M+H]⁺; ¹H NMR (400 MHz, CDCl₃) δ ppm 1.29 (t, J=7.2 Hz, 3H),2.05-2.14 (m, 1H), 2.50 (dd, J=16.8, 11.2 Hz, 1H), 2.68-2.86 (m, 4H),3.48-3.58 (m, 1H), 4.16-4.24 (m, 2H), 6.66 (dd, J=8.6, 2.4 Hz, 1H), 6.85(d, J=2.4 Hz, 1H), 7.15 (d, J=8.7 Hz, 1H), 8.4 (s, 1H).

Step E: Preparation of 3-Cyano-5-(trifluoromethoxy)benzoyl Chloride

Oxalyl chloride (2.0 M in DCM, 0.636 mL, 1.272 mmol) was added to neat3-cyano-5-(trifluoromethoxy)benzoic acid (98 mg, 0.424 mmol) and onedrop of DMF was added. The reaction mixture was stirred at roomtemperature for 30 min and then concentrated under reduced pressure.

Step F: Preparation of3-(Hydroxymethyl)-5-(trifluoromethoxy)benzonitrile

3-Cyano-5-(trifluoromethoxy)benzoyl chloride (844 mg, 3.38 mmol) wasdissolved in THF (10 mL) and cooled to 0° C. Sodium borohydride (320 mg,8.45 mmol) was added, followed by methanol (2 mL) and the reaction wasstirred for 20 min at 0° C. before it was allowed to warm to roomtemperature. After 2 h, the reaction mixture was acidified to pH 3 with1.0 M HCl. The aqueous mixture was extracted with ethyl acetate and thecombined extracts were dried over sodium sulfate, filtered andconcentrated under reduced pressure. The residue was purified by silicagel column chromatography to give the title compound (440 mg). LCMSm/z=218.3 [M+H]⁺.

Step G: Preparation of2-(7-(3-Cyano-5-(trifluoromethoxy)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)aceticAcid

Ethyl 2-(7-hydroxy-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetate(100 mg, 0.386 mmol) and3-(hydroxymethyl)-5-(trifluoromethoxy)benzonitrile (84 mg, 0.386 mmol)were dissolved in THF (3.0 mL) and cooled to 0° C. Triphenylphosphine(202 mg, 0.771 mmol) and diisopropylazodicarboxylate (DIAD) (0.15 mL,0.771 mmol) were added. The mixture was warmed to room temperature andstirred for 1 h. Additional DIAD (0.15 mL, 0.771 mmol) andtriphenylphosphine (202 mg, 0.771 mmol) were added and the reactionmixture was stirred for 1 h. The reaction mixture was diluted with waterand extracted three times with ethyl acetate. The combined organics weredried over sodium sulfate, filtered and concentrated under reducedpressure. The residue was purified by silica gel column chromatographyto give 50.8 mg of impure ethyl2-(7-(3-cyano-5-(trifluoromethoxy)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetate.The material was dissolved in dioxane (1.3 mL) and 1.0 M aqueous LiOH(0.33 mL, 0.33 mmol) was added. The reaction was monitored by HPLC untiljudged complete and then acidified to pH 2 with 1.0 M HCL. The aqueousmixture was extracted three times with ethyl acetate. The combinedorganics were dried over sodium sulfate, filtered and concentrated underreduced pressure. The residue was purified by silica gel columnchromatography followed by HPLC to give the title compound (1.1 mg).LCMS m/z=431.2 [M+H]⁺; ¹H NMR (400 MHz, CD₃OD) δ ppm 2.11-2.20 (m, 1H),2.50 (dd, J=15.8, 8.0 Hz, 1H), 2.66-2.84 (m, 4H), 3.51-3.60 (m, 1H),5.18 (s, 2H), 6.78 (dd, J=8.8, 2.5 Hz, 1H), 6.95 (d, J=2.4 Hz, 1H), 7.20(d, J=8.9 Hz, 1H), 7.64 (s, 1H), 7.73 (s, 1H), 7.84 (s, 1H).

Example 1.2 Preparation of2-(7-(3,5-Bis(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)aceticAcid (Compound 10) Step A: Preparation of Ethyl2-(7-(3,5-Bis(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetate

Ethyl 2-(7-hydroxy-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetate (61mg, 0.235 mmol), was dissolved in DMF (1.0 mL) and cesium carbonate (77mg, 0.235 mmol) and 1-(bromomethyl)-3,5-bis(trifluoromethyl)benzene (72mg, 0.235 mmol) were added. The reaction mixture was stirred at roomtemperature for 16 h and then filtered through a pad of Celite®. Thefiltrate was diluted with water and extracted three times with ethylacetate. The combined organics were dried over sodium sulfate, filteredand concentrated under reduced pressure. The residue was purified bysilica gel column chromatography to give the title compound (28.6 mg).LCMS m/z=486.4 [M+H]⁺.

Step B: Preparation of2-(7-(3,5-Bis(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)aceticAcid

Ethyl2-(7-(3,5-bis(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetate(28.6 mg, 0.059 mmol) was dissolved in dioxane (1.0 mL) and 1.0 Maqueous LiOH (0.166 mL, 0.166 mmol) was added. The solution was stirredat room temperature for 3 h before it was acidified to pH 3 with 1.0 MHCl and extracted twice with ethyl acetate. The combined extracts weredried over sodium sulfate, filtered and concentrated under reducedpressure to give the title compound (23 mg). LCMS m/z=458.3 [M+H]⁺; ¹HNMR (400 MHz, CDCl₃) δ ppm 2.12-2.24 (m, 1H), 2.61 (dd, J=17.0, 10.7 Hz,1H), 2.73-2.89 (m, 4H), 3.53-3.63 (m, 1H), 5.19 (s, 2H), 6.86 (dd,J=8.6, 2.5 Hz, 1H), 7.0 (d, J=2.5 Hz, 1H), 7.24 (d, J=8.8 Hz, 1H), 7.82(s, 1H), 7.94 (s, 2H), 8.33 (s, 1H).

Example 1.3 Preparation of2-(7-(3-Cyano-4-isopropoxybenzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)aceticAcid (Compound 5) Step A: Preparation of5-(Hydroxymethyl)-2-isopropoxybenzonitrile

From 3-cyano-4-isopropoxybenzoic acid, in a similar manner to the onedescribed in Example 1.1, Step E and F, the title compound was obtained.¹H NMR (400 MHz, CDCl₃) δ ppm 1.40 (d, J=6.2 Hz, 6H), 1.72 (t, J=5.6 Hz,1H), 4.6-4.69 (m, 3H), 6.95 (d, J=8.8 Hz, 1H), 7.50 (dd, J=8.6, 2.0 Hz,1H), 7.55 (d, J=2.3 Hz, 1H).

Step B: Preparation of 5-(Chloromethyl)-2-isopropoxybenzonitrile

5-(Hydroxymethyl)-2-isopropoxybenzonitrile (5.96 g, 31.2 mmol) wasdissolved in toluene (90 mL) and thionyl chloride (13.65 mL, 187 mmol)was added. The reaction mixture was warmed to 75° C. and stirred for 20min. The reaction mixture was diluted with hexanes and washed with waterand saturated aqueous sodium bicarbonate. The hexane solution was driedover sodium sulfate, filtered and concentrated under reduced pressure togive the title compound (5.6 g). ¹H NMR (400 MHz, CDCl₃) δ ppm 1.41 (d,J=6.1 Hz, 6H), 4.52 (s, 2H), 4.66 (septet, J=6.1 Hz, 1H), 6.95 (d, J=8.6Hz, 1H), 7.51 (dd, J=8.7, 2.4 Hz, 1H), 7.57 (d, J=2.3 Hz, 1H).

Step C: Preparation of Ethyl2-(7-(3-Cyano-4-isopropoxybenzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetate

Ethyl 2-(7-hydroxy-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetate(1.237 g, 4.77 mmol) was dissolved in DMF (12 mL) and cesium carbonate(1.554 g, 4.77 mmol) was added. The reaction mixture was stirred at roomtemperature for 10 min and 5-(chloromethyl)-2-isopropoxybenzonitrile(1.0 g, 4.77 mmol) was added. The reaction mixture was stirred at 40° C.for 2 h before it was cooled to room temperature. The heterogenousmixture was filtered through Celite® and the filtrate was concentratedunder reduced pressure. The residue was purified by silica gel columnchromatography to give the title compound (1.32 g). LCMS m/z=433.5[M+H]⁺; ¹H NMR (400 MHz, CDCl₃) δ ppm 1.29 (t, J=7.2 Hz, 3H), 1.40 (d,J=6.1 Hz, 6H), 2.05-2.16 (m, 1H), 2.50 (dd, J=16.7, 11.1 Hz, 1H),2.69-2.88 (m, 4H), 3.50-3.59 (m, 1H), 4.16-4.26 (m, 2H), 4.65 (septet,J=6.1 Hz, 1H), 5.00 (s, 2H), 6.80 (dd, J=8.7, 2.5 Hz, 1H), 6.94-6.97 (m,2H), 7.20 (d, J=8.8 Hz, 1H), 7.58 (dd, J=8.7, 2.3 Hz, 1H), 7.64 (d,J=2.0 Hz, 1H), 8.45 (s, 1H).

Step D: Preparation of2-(7-(3-Cyano-4-isopropoxybenzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)aceticAcid

Ethyl2-(7-(3-cyano-4-isopropoxybenzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetate(1.32 g, 3.05 mmol) was dissolved in dioxane (34 mL) and 1.0 M aqueousLiOH (9.16 mL, 9.16 mmol) was added. The reaction was stirred at roomtemperature for 6 h and then warmed to 35° C. and stirred for oneadditional hour. After cooling to room temperature, the reaction wasacidified to pH 3 with 1.0 M HCl and partitioned between water and ethylacetate. The organics were removed and the aqueous layer was extractedtwice with ethyl acetate. The combined extracts were dried over sodiumsulfate, filtered and concentrated to give the title compound (1.23 g).LCMS m/z=405.6 [M+H]⁺; ¹H NMR (400 MHz, DMSO-d₆) δ ppm 1.32 (d, J=5.9Hz, 6H), 2.03-2.13 (m, 1H), 2.35 (dd, J=15.9, 9.0 Hz, 1H), 2.58-2.77 (m,4H), 3.41-3.51 (m, 1H), 4.79 (septet, J=5.9 Hz, 1H), 5.01 (s, 2H), 6.69(dd, J=8.8, 2.4 Hz, 1H), 6.91 (d, J=2.4 Hz, 1H), 7.19 (d, J=8.6 Hz, 1H),7.28 (d, J=8.8 Hz, 1H), 7.70 (dd, J=8.8, 2.3 Hz, 1H), 7.76 (d, J=2.1 Hz,1H), 10.45 (s, 1H), 12.1 (bs, 1H).

Resolution Via Chiral HPLC.

Column: normal phase preparative ChiralCel OD, 50×500 mm ID, 20 μmparticle size

Eluent: 75% Hexane/25% Isopropanol, with 0.05% trifluoroacetic acid

Gradient: Isocratic

Flow: 60 mL/min

Detector: 254 nm

Retention Times: 1^(st) enantiomer: 33 min; 2^(nd) enantiomer: 40 min.

Example 1.4 Preparation of2-(7-(4-Cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)aceticAcid (Compound 12) Step A: Preparation of Methyl4-Chloro-3-(trifluoromethyl)benzoate

To a solution of 4-chloro-3-(trifluoromethyl)benzoic acid (10.37 g, 46.2mmol) in methanol (100 mL) was added concentrated sulfuric acid (0.51mL, 9.24 mmol). The mixture was heated under reflux overnight. Themixture was allowed to cool to room temperature and concentrated underreduced pressure to form a solid. The solid was filtered and washed withwater. The solid was then stirred with saturated aqueous sodiumbicarbonate solution to remove any residual sulfuric acid, filtered anddried under vacuum to give the title compound as a white solid (10.18g). ¹H NMR (400 MHz, CDCl₃) δ ppm 3.96 (s, 3H), 7.60 (d, J=8.34 Hz, 1H),8.14 (dd, J=8.34, 2.02 Hz, 1H), 8.37 (d, J=2.02 Hz, 1H).

Step B: Preparation of Methyl 4-Cyclopentyl-3-(trifluoromethyl)benzoate

To zinc(II) chloride (0.5 M solution in tetrahydrofuran, 88.0 mL, 44.0mmol) was added cyclopentylmagnesium chloride (2 M solution in ether,20.5 mL, 41.1 mmol). The resulting suspension was stirred at roomtemperature for 1 h. To the above suspension was added methyl4-chloro-3-(trifluoromethyl)benzoate (7.00 g, 29.3 mmol) andbis(tri-tert-butylphosphine)palladium (1.35 g, 2.64 mmol) at roomtemperature. The mixture was heated under reflux for 2 h. The mixturewas allowed to cool to room temperature, quenched with saturated aqueoussodium bicarbonate solution and filtered. The filtrate was extractedwith ethyl acetate. The organic layer was dried over anhydrous sodiumsulfate, concentrated under reduced pressure and purified by silica gelcolumn chromatography to give the title compound as an oil (7.64 g).LCMS m/z=273.2 [M+H]⁺; ¹H NMR (400 MHz, CDCl₃) δ ppm 1.57-1.66 (m, 2H),1.68-1.82 (m, 2H), 1.82-1.94 (m, 2H), 2.04-2.21 (m, 2H), 3.33-3.49 (m,1H), 3.93 (s, 3H), 7.54 (d, J=8.21 Hz, 1H), 8.13 (dd, J=8.34, 1.77 Hz,1H), 8.27 (s, 1H).

Step C: Preparation of (4-Cyclopentyl-3-(trifluoromethyl)phenyl)methanol

To a solution of methyl 4-cyclopentyl-3-(trifluoromethyl)benzoate (8.16g, 30.0 mmol) in 1,4-dioxane (200 mL) was added lithium borohydridesolution (2 M in tetrahydrofuran, 30.0 mL, 59.9 mmol). The mixture washeated under reflux for 2.5 h. The mixture was allowed to cool to roomtemperature and carefully quenched with 1 N aqueous HCl solution to pH5. The organic layer was separated and the aqueous layer was extractedwith ethyl acetate. The combined organic layers were dried overanhydrous sodium sulfate, concentrated under reduced pressure andpurified by silica gel column chromatography to give the title compoundas a colorless oil (1.21 g). ¹H NMR (400 MHz, CDCl₃) δ ppm 1.56-1.63 (m,2H), 1.66-1.77 (m, 2H), 1.81-1.91 (m, 2H), 2.03-2.15 (m, 2H), 3.37(quintet, J=8.00 Hz, 1H), 4.71 (d, J=4.29 Hz, 2H), 7.45-7.47 (m, 1H),7.49 (d, J=1.14 Hz, 1H), 7.60 (s, 1H).

Step D: Preparation of4-(Chloromethyl)-1-cyclopentyl-2-(trifluoromethyl)benzene

To (4-cyclopentyl-3-(trifluoromethyl)phenyl)methanol (1.21 g, 4.95 mmol)was added thionyl chloride (5.5 mL, 74.2 mmol). The mixture was heatedat 50° C. for 2 h before it was allowed to cool to room temperature andstirred at room temperature overnight. The mixture was poured into anice and stirred for 5 min before it was extracted with dichloromethane.The organic extract was washed with saturated aqueous sodium bicarbonatesolution, dried over anhydrous sodium sulfate and concentrated underreduced pressure to give the title compound as an oil (1.16 g). ¹H NMR(400 MHz, CDCl₃) δ ppm 1.55-1.63 (m, 2H), 1.69-1.77 (m, 2H), 1.82-1.90(m, 2H), 2.05-2.13 (m, 2H), 3.37 (quintet, J=8.59 Hz, 1H), 4.58 (s, 2H),7.46 (d, J=8.00 Hz, 1H), 7.52 (d, J=8.00 Hz, 1H), 7.61 (d, J=1.52 Hz,1H).

Step E: Preparation of Ethyl2-(7-(4-Cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetate

To a solution of ethyl2-(7-hydroxy-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetate (50.0 mg,0.193 mmol) and4-(chloromethyl)-1-cyclopentyl-2-(trifluoromethyl)benzene (152.0 mg,0.578 mmol) in DMF (3 mL) was added cesium carbonate (75.0 mg, 0.231mmol). The mixture was stirred at room temperature overnight, filteredthrough Celite®, and concentrated under reduced pressure. The residuewas purified by HPLC to give the title compound as a light pink oil(38.7 mg). LCMS m/z=486.5 [M+H]⁺.

Step F: Preparation of2-(7-(4-Cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)aceticAcid

To a solution of ethyl2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetate(38.7 mg, 0.080 mmol) in a mixed solvent of methanol (1.5 mL),tetrahydrofuran (0.5 mL), and water (0.5 mL) was added LiOH hydrate(11.7 mg, 0.279 mmol). The mixture was stirred at room temperatureovernight before the mixture was acidified to pH 4 with 1 N aqueous HClsolution and extracted with ethyl acetate. The organic layer was driedover anhydrous sodium sulfate, concentrated under reduced pressure anddried under vacuum. The foam was triturated with water to give a solid.The solid was filtered to give the title compound as a light pink solid(25.7 mg). LCMS m/z=458.4 [M+H]⁺; ¹H NMR (400 MHz, DMSO-d₆) δ ppm1.56-1.70 (m, 4H), 1.80-1.87 (m, 2H), 1.95-2.11 (m, 3H), 2.34 (dd,J=16.04, 8.97 Hz, 1H), 2.59-2.74 (m, 4H), 3.21-3.25 (m, 1H), 3.41-3.49(m, 1H), 5.11 (s, 2H), 6.70 (dd, J=8.72, 2.40 Hz, 1H), 6.92 (d, J=2.27Hz, 1H), 7.19 (d, J=8.72 Hz, 1H), 7.61 (d, J=8.00 Hz, 1H), 7.68 (d,J=8.00 Hz, 1H), 7.70 (s, 1H), 10.45 (s, 1H), 12.18 (bs, 1H).

Resolution Via Chiral HPLC.

Column: normal phase preparative ChiralCel OD, 50×500 mm ID, 20 μmparticle size

Eluent: IPA containing 0.05% TFA/hexanes containing 0.05% TFA (8/92)

Gradient: Isocratic

Flow: 60 mL/min

Detector: 220 nm

Retention Times: 1^(st) enantiomer: 38.9 min; 2^(nd) enantiomer: 48.4min.

Example 1.5 Preparation of2-(7-((5-Isopropoxypyrazin-2-yl)methoxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)aceticAcid (Compound 11) Step A: Preparation of 2-Isopropoxy-5-methylpyrazine

To a solution of 2-bromo-5-methylpyrazine (3 g, 17.34 mmol) in2-propanol (14 mL) was added sodium propan-2-olate (3.56 g, 43.3 mmol)and heated under microwave irradiation at 115° C. for 1.1 h. The organicsolvent was evaporated before water was added to the residue. Themixture was extracted with dichloromethane (2×75 mL). The organic phasewas dried over sodium sulfate, filtered and concentrated. The residuewas purified by silica gel column chromatography to give the titlecompound as a brown oil (1.0 g). LCMS m/z=153.4 [M+H]⁺; ¹H NMR (400 MHz,CDCl₃) δ ppm 1.27 (d, J=6.19 Hz, 6H), 2.39 (s, 3H), 5.10-5.20 (m, 1H),7.84 (s, 1H), 7.99 (s, 1H).

Step B: Preparation of 2-Isopropoxy-5-methylpyrazine

A mixture of 2-isopropoxy-5-methylpyrazine (0.250 g, 1.65 mmol), NBS(0.293 g, 1.65 mmol) and AIBN (0.270 g, 1.65 mmol) in toluene (5 mL) wasrefluxed for 1 h after which 1.0 eq of NBS was added. The reactionmixture was heated under reflux for 20 min before it was cooled to roomtemperature. The solids were removed by filtration and the filtrate wasconcentrated under vacuum. The residue was purified by silica gel columnchromatography to give the title compound as a brown oil (35 mg).

Step C: Preparation of 2-(7-((5-Isopropoxypyrazin-2-yl)methoxy)-1,2,3,4tetrahydrocyclopenta[b]indol-3-yl)acetic Acid

Ethyl 2-(7-hydroxy-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetate(0.035 g, 0.135 mmol) and cesium carbonate (0.048 g, 0.148 mmol) weredissolved in DMF (0.5 mL) and stirred at room temperature for 5 min. Tothis mixture at 0° C. was added a solution of2-(bromomethyl)-5-isopropxypyrazine (0.034 g, 0.148 mmol) in DMF (0.20mL) and was stirred at room temperature for 60 h. The solids wereremoved by filtration. The filtrate was purified by HPLC to give ethyl2-(7-((5-isopropoxypyrazin-2-yl)methoxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetate(11 mg). To the ester dissolved in dioxane (288 μL) was added aqueousLiOH (1 N, 72 μL). The mixture was stirred at room temperature for 16 h;before more aqueous LiOH (1 N, 200 μL) was added. Stirring was continuedfor 1 h. To the reaction mixture was added water (1.5 mL) and thereaction mixture was acidified to pH 3 with 1 N HCl. The mixture waspurified by HPLC to give the title compound as a yellow solid (7 mg).LCMS m/z=382.4 [M+H]⁺; ¹H NMR (500 MHz, CDCl₃) δ ppm 1.32 (d, J=6.31 Hz,6H), 1.95-2.18 (m, 1H), 2.28-2.41 (m, 1H), 2.56-2.79 (m, 4H), 3.36-3.56(m, 1H), 5.09 (s, 2H), 5.15-5.33 (m, 1H), 6.71 (d, J=8.83 Hz, 1H), 6.94(s, 1H), 7.20 (d, J=8.83 Hz, 1H), 8.20 (s, 1H), 8.30 (s, 1H), 10.37 (s,1H).

Example 1.6 Preparation of2-(7-(3-Cyano-4-(trifluoromethoxy)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)aceticAcid (Compound 7) Step A: Preparation of3-Bromo-4-(trifluoromethoxy)benzoic Acid

4-(Trifluoromethoxy)benzoic acid (2 g, 9.70 mmol) and iron(III) chloride(1.574 g, 9.70 mmol) were suspended in nitromethane (20 mL). To thismixture was added bromine (0.497 mL, 9.70 mmol) at 0° C. The solutionwas heated under microwave irradiation at 110° C. for 2 h. The reactionmixture was added to cold water (100 mL) and extracted with ethylacetate(2×100 mL). The organic phase was washed with an aqueous solution ofsodium thiosulfate pentahydrate and brine. The organic layer wasconcentrated and the residue was purified by HPLC to give the titlecompound as a white solid (1.5 g). LCMS m/z=287.0 [M+H]⁺; ¹H NMR (400MHz, DMSO-d₆) δ ppm 7.67 (d, 1H), 8.06 (d, 1H), 8.29 (s, 1H), 13.47 (s,1H).

Step B: Preparation of 3-Cyano-4-(trifluoromethoxy)benzoic Acid

3-Bromo-4-(trifluoromethoxy)benzoic acid (1.4 g, 4.91 mmol) andcyanocopper (0.572 g, 6.39 mmol) were mixed in N-methyl-2-pyrrolidinone(NMP) (14 mL). The mixture was heated in a microwave at 200° C. for 2 h.The reaction was diluted with dichloromethane (150 mL). Celite® wasadded and the mixture was stirred vigorously for 10 min. The solids wereremoved by filtration. The organic layer was washed with water (125 mL)and concentrated. The residue was purified by HPLC to give the titlecompound as an off-white solid (0.979 g). LCMS m/z=232.3 [M+H]⁺.

Step C: Preparation of5-(Hydroxymethyl)-2-(trifluoromethoxy)benzonitrile

From 3-cyano-4-(trifluoromethoxy)benzoic acid, in a similar manner tothe one described in Example 1.3, Step A, the title compound wasobtained as a clear oil. LCMS m/z=218.2 [M+H]⁺. ¹H NMR (400 MHz, CDCl₃)δ ppm 4.81 (s, 2H), 7.40-7.44 (m, 1H), 7.67-7.71 (m, 2H), 7.78 (s, 1H).

Step D: Preparation of 5-(Chloromethyl)-2-(trifluoromethoxy)benzonitrile

5-(Hydroxymethyl)-2-(trifluoromethoxy)benzonitrile (0.150 g, 0.691 mmol)was taken up in toluene (2 mL) and thionyl chloride (0.303 mL, 4.14mmol) was added. The mixture was heated at 75° C. for 15 min. Water wasadded and the mixture was extracted with hexanes (2×75 mL). The organicswere treated with aqueous NaHCO₃. The organic layer was separated, driedover magnesium sulfate, filtered and concentrated to give the titlecompound as a colorless oil (120 mg). LCMS m/z=236.2 [M+H]⁺. ¹H NMR (500MHz, CDCl₃) δ ppm 1.98 (s, 1H), 4.52 (s, 2H), 7.32-7.34 (m, 1H),7.59-7.62 (m, 1H), 7.68 (s, 1H).

Step E: Preparation of2-(7-(3-Cyano-4-(trifluoromethoxy)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)aceticAcid

Ethyl 2-(7-hydroxy-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetate(0.100 g, 0.386 mmol) and cesium carbonate (0.138 g, 0.424 mmol) weredissolved in DMF (1.0 mL), stirred at room temperature for 10 min,followed by addition of5-(chloromethyl)-2-(trifluoromethoxy)benzonitrile (0.100 g, 0.424 mmol)in DMF (0.300 mL) at 0° C. This mixture was stirred at room temperaturefor 3 h. The reaction mixture was quenched with water and extracted withethylacetate (2×50 mL). The organic phase was washed with brine andconcentrated. The residue was taken up in dioxane (4 mL) before aqueous1 N LiOH (1.3 mL) was added. The mixture was stirred at room temperaturefor 2.5 h before it was quenched with water and acidified to pH 3 usingaqueous 3 N HCl. The mixture was purified by HPLC to give the titlecompound (0.040 g). LCMS m/z=431.2 [M+H]⁺, ¹H NMR (500 MHz, DMSO-d₆) δppm 2.00-2.20 (m, 1H), 2.01-2.21 (m, 1H), 2.23-2.43 (m, 1H), 2.55-2.83(m, 4H), 3.33-3.57 (m, 1H), 5.16 (s, 2H), 6.73 (dd, J=8.83, 2.52 Hz,1H), 6.94 (s, 1H), 7.21 (d, J=8.83 Hz, 1H), 7.69 (dd, J=8.67, 1.42 Hz,1H), 7.94 (dd, J=8.67, 2.05 Hz, 1H), 8.09 (s, 1H), 10.40 (s, 1H).

Example 1.7 Preparation of2-(7-(2,4-Bis(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)aceticAcid (Compound 8) Step A: Preparation of Ethyl2-(7-(2,4-Bis(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetate

To a mixture of ethyl2-(7-hydroxy-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetate (0.130 g,0.5 mmol) and K₂CO₃ (0.069 g, 0.500 mmol) in DMF (1 mL) was added1-(bromomethyl)-2,4-bis(trifluoromethyl)benzene (0.154 g, 0.500 mmol).The mixture was heated at 70° C. overnight, taken up in EtOAc, washedwith water (thrice) and brine. The organics were dried over MgSO₄ andconcentrated. The residue was purified by silica gel columnchromatography to give the title compound an orange solid (0.195 g).LCMS m/z 486.3 [M+H]⁺.

Step B: Preparation of2-(7-(2,4-Bis(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)aceticAcid

To a solution of ethyl2-(7-(2,4-bis(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetate(0.191 g, 0.393 mmol) in dioxane (1.312 mL) and water (0.656 mL) wasadded NaOH (0.826 mL, 0.826 mmol). The mixture was heated under refluxfor 2 h, cooled to room temperature and diluted with water. Afterwashing with DCM, the aqueous layer was acidified with 1 M HCl andextracted with EtOAc (thrice). The combined extracts were washed withbrine, dried over MgSO₄ and concentrated to give the title compound as amagenta solid (19.9 mg). LCMS m/z=458.1 [M+H]⁺, ¹H NMR (400 MHz,DMSO-d₆) δ ppm 2.03-2.13 (m, 1H), 2.35 (dd, J=15.98, 9.03 Hz, 1H),2.58-2.78 (m, 4H), 3.41-3.52 (m, 1H), 5.31 (s, 2H), 6.72 (dd, J=8.78,2.46 Hz, 1H), 6.92 (d, J=2.40 Hz, 1H), 7.23 (d, J=8.72 Hz, 1H),7.99-8.19 (m, 3H), 10.52 (s, 1H), 12.19 (s, 1H).

Example 1.8 Preparation of2-(7-(4-Cyclohexyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)aceticAcid (Compound 2) Step A: Preparation of1-Chloro-4-(chloromethyl)-2-(trifluoromethyl)benzene

(4-Chloro-3-(trifluoromethyl)phenyl)methanol (5.1 g, 24.22 mmol) wasadded in small portions to thionyl chloride (20 mL, 275 mmol). Thereaction mixture was stirred at 50° C. for 18 h and heated under refluxfor 23 h. The mixture was concentrated and dried under high vacuum togive the title compound as a colorless liquid (5.41 g). ¹H NMR (400 MHz,CDCl₃) δ ppm 4.58 (s, 2H), 7.50-7.51 (m, 2H), 7.71 (s, 1H).

Step B: Preparation of Ethyl2-(7-(4-Chloro-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetate

A mixture of ethyl2-(7-hydroxy-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetate (222 mg,0.856 mmol), 1-chloro-4-(chloromethyl)-2-(trifluoromethyl)benzene (240mg, 1.048 mmol), and cesium carbonate (165 mg, 0.856 mmol) in DMF (5 mL)was stirred at room temperature. After 3 days, more cesium carbonate(165 mg, 0.856 mmol) was added. After stirring for an additional 2 d,the mixture was extracted with water and CH₂Cl₂. The organics were driedover MgSO₄, filtered and concentrated. The residue was purified bysilica gel column chromatography to give the title compound as a whitesolid (258 mg). LCMS m/z=452.1 [M+H]⁺; ¹H NMR (400 MHz, CDCl₃) δ ppm1.29 (t, J=7.2 Hz, 3H), 2.06-2.14 (m, 1H), 2.47-2.54 (m, 1H), 2.71-2.86(m, 4H), 3.51-3.57 (m, 1H), 4.17-4.25 (m, 2H), 5.10 (s, 2H), 6.82 (dd,J=8.8, 2.5, 1H), 6.96 (d, J=2.5 Hz, 1H), 7.21 (dd, J=8.8, 0.32 Hz, 1H),7.49-7.52 (m, 1H), 7.58 (dd, J=8.2, 2.6 Hz, 1H), 7.80 (d, J=1.92 Hz,1H), 8.48 (s, 1H).

Step C: Preparation of2-(7-(4-Cyclohexyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)aceticAcid

A mixture of ethyl2-(7-(4-chloro-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetate(50 mg, 0.111 mmol), 0.5 M cyclohexylzinc(II) bromide (0.5 M in THF, 3mL, 1.5 mmol), and bis(tri-t-butylphosphine)palladium (3 mg, 5.87 μmol)were stirred under reflux for 18 h. The mixture was allowed to cool toroom temperature before water (1 mL), MeOH (1 mL) and LiOH hydrate (70mg, 1.668 mmol) were added. The mixture was stirred at room temperaturefor 2 h. The mixture was purified by HPLC. Fractions containing productwere basified with 1 M NaHCO₃ and partially concentrated. The residuewas extracted with 0.5 M citric acid and CH₂Cl₂. The organics were driedover MgSO₄, filtered and concentrated to give the title compound as atanned sticky solid (14.4 mg). LCMS m/z=472.2 [M+H]⁺; ¹H NMR (400 MHz,CDCl₃) δ ppm 1.38-1.50 (m, 4H), 1.76-1.85 (m, 6H), 2.11-2.17 (m, 1H),2.58-2.65 (m, 1H), 2.75-2.93 (m, 5H), 3.56-3.60 (m, 1H), 5.07 (s, 2H),6.84 (dd, J=8.8, 2.5 Hz, 1H), 7.00 (d, J=2.4 Hz, 1H), 7.21 (d, J=8.8 Hz,1H), 7.46 (d, J=8.1 Hz, 1H), 7.59 (d, J=8.1 Hz, 1H), 7.70 (d, J=1.2 Hz,1H), 8.27 (s, 1H).

Example 1.9 Preparation of2-(7-(4-(Pyrrolidin-1-yl)-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)aceticAcid (Compound 13) Step A: Preparation of Ethyl2-(7-(4-(Pyrrolidin-1-yl)-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetate

A mixture of ethyl2-(7-(4-chloro-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetate(50.9 mg, 0.113 mmol), pyrrolidine (0.047 mL, 0.563 mmol),diacetoxypalladium (1.264 mg, 5.63 μmol),biphenyl-2-yl-di-tert-butylphosphine (3.36 mg, 11.0 μmol) and sodium2-methylpropan-2-olate (27.1 mg, 0.282 mmol) in dioxane (3 mL) washeated under microwave irradiation at 120° C. for 2 h. The mixture waspurified by HPLC. Fractions containing product were basified with 1 MNaHCO₃ and concentrated. The residue was extracted with 0.5 M citricacid and CH₂Cl₂. The organics were dried over MgSO₄, filtered, andconcentrated to give the title compound as a white solid (17.8 mg). LCMSnt/z=487.4 [M+H]⁺.

Step B: Preparation of2-(7-(4-(Pyrrolidin-1-yl)-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)aceticAcid

To a solution of ethyl2-(7-(4-(pyrrolidin-1-yl)-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetate(17.8 mg, 0.037 mmol) in 5 mL (THF/water/MeOH 3:1:1), LiOH hydrate (7.68mg, 0.183 mmol) was added. After stirring at room temperature for 2 h,the mixture was partially concentrated and the residue was extractedwith 0.5 M citric acid and CH₂Cl₂. The organics were dried over MgSO₄,filtered and concentrated to give the title compound as a brownishsticky solid (16.3 mg). LCMS m/z=459.4 [M+H]⁺; ¹H NMR (400 MHz, CDCl₃) δppm 1.83-1.95 (m, 4H), 2.09-2.16 (m, 1H), 2.57-2.64 (m, 1H), 2.73-2.90(m, 4H), 3.27-3.35 (m, 4H), 3.53-3.59 (m, 1H), 4.99 (s, 2H), 6.82 (dd,J=8.8, 2.4 Hz, 1H), 6.96-7.01 (m, 2H), 7.19 (d, J=8.7 Hz, 1H), 7.45 (dd,J=8.6 Hz, 2.0 Hz, 1H), 7.66 (d, J=2.0 Hz, 1H), 8.27 (s, 1H).

Example 1.10 Preparation of2-(7-(4-Isobutyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)aceticAcid (Compound 14)

Ethyl2-(7-(4-chloro-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetate(200 mg, 0.443 mmol) was dissolved in THF (7 mL) and isobutylzinc(II)bromide (2.66 mL, 1.328 mmol) and bis(tri-t-butylphosphine)palladium(0)(0.011 g, 0.022 mmol) were added. The reaction was stirred at roomtemperature for 16 h and warmed to 50° C. After stirring for 24 h,isobutylzinc(II) bromide (4 mL) was added and the mixture was heated to90° C. The reaction was cooled to room temperature, and 1.0 M LiOH (5mL) and dioxane (5 mL) were added. The reaction was stirred at roomtemperature for 24 h and then acidified to pH 3 with 1 M HCl. Theaqueous mixture was extracted three times with EtOAc. The combinedextracts were dried over sodium sulfate, filtered and concentrated underreduced pressure. The residue was purified by silica gel columnchromatography to provide the title compound (33 mg). LCMS m/z=446.7[M+H]⁺; ¹H NMR (400 MHz, DMSO-d₆) δ ppm 0.89 (d, J=6.6 Hz, 6H),1.87-1.98 (m, 1H), 2.03-2.13 (m, 1H), 2.35 (dd, J=15.9, 9.0 Hz, 1H),2.60-2.76 (m, 6H), 3.42-3.50 (m, 1H), 5.12 (s, 2H), 6.71 (dd, J=8.6, 2.4Hz, 1H), 6.93 (d, J=2.4 Hz, 1H), 7.20 (d, J=8.7 Hz, 1H), 7.47 (d, J=8.0Hz, 1H), 7.67 (d, J=8.2 Hz, 1H), 7.75 (d, J=1.4 Hz, 1H), 10.47 (bs, 1H).

Example 1.11 Preparation of2-(7-(4-Neopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)aceticAcid (Compound 15)

Ethyl2-(7-(4-chloro-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetate(200 mg, 0.443 mmol) was dissolved in THF (7.0 mL) and neopentylzinc(II)iodide (2.66 mL of a 0.5 M solution in THF) andbis(tri-t-butylphosphine)palladium(0) (0.011 g, 0.022 mmol) were added.The reaction mixture was stirred at room temperature for 16 h and thenwarmed to 50° C. After stirring for 24 h, neopentylzinc(II) iodide (5.0mL of a 0.5 M solution in THF) was added and the reaction mixture washeated to 90° C. The reaction vessel was cooled to room temperaturebefore 1.0 M LiOH (5 mL) and dioxane (5.0 mL) were added. After stirringfor 24 h, the reaction was acidified with 1.0 M HCl to pH 3 andextracted three times with EtOAc. The combined extracts were dried oversodium sulfate, filtered and concentrated under reduced pressure. Theresidue was purified by HPLC. The purified fractions were neutralizedwith sodium bicarbonate and then acidified to pH 5 with 1.0 M citricacid. The aqueous mixture was extracted with EtOAc and the organic layerwas washed two times with water. The EtOAc layer was dried over sodiumsulfate, filtered and concentrated under reduced pressure to provide thetitle compound (12.3 mg). LCMS m/z=460.6 [M+H]⁺; ¹H NMR (400 MHz, CDCl₃)δ ppm 0.96 (s, 9H), 2.08-2.19 (m, 1H), 2.61 (dd, J=17.0, 10.9 Hz, 1H),2.73-2.90 (m, 6H), 3.54-3.63 (m, 1H), 5.09 (s, 2H), 6.85 (dd, J=8.8, 2.5Hz, 1H), 7.0 (d, J=2.5 Hz, 1H), 7.22 (d, J=9.0 Hz, 1H), 7.36 (d, J=8.0Hz, 1H), 7.55 (d, J=8.1 Hz, 1H), 7.74 (d, J=1.1 Hz, 1H), 8.29 (bs, 1H).

Example 1.12 Preparation of2-(7-(4-Chloro-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)aceticAcid (Compound 16)

The title compound was isolated as a by-product from Example 1.11. LCMSm/z=424.2 [M+H]⁺; ¹H NMR (400 MHz, CDCl₃) δ ppm 2.09-2.19 (m, 1H), 2.61(dd, J=17.3, 11.0 Hz, 1H), 2.72-2.89 (m, 4H), 3.53-3.63 (m, 1H), 5.10(s, 2H), 6.83 (dd, J=8.7, 2.4 Hz, 1H), 6.97 (d, J=2.4 Hz, 1H), 7.22 (d,J=8.6 Hz, 1H), 7.50 (d, J=8.2 Hz, 1H), 7.57 (dd, J=8.2, 1.6 Hz, 1H),7.80 (d, J=1.8 Hz, 1H), 8.33 (bs, 1H).

Example 1.13 Preparation of2-(7-(3-Cyano-4-cyclohexylbenzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)aceticAcid (Compound 17) Step A: Preparation of Methyl3-cyano-4-hydroxybenzoate

To a mixture of methyl 3-bromo-4-hydroxybenzoate (1.78 g, 7.70 mmol) andcopper(I) cyanide (0.897 g, 10.02 mmol) was added NMP (10 mL). Themixture was heated to 200° C. for 2 h under microwave irradiation. Themixture was diluted with ethyl acetate and quenched with 1 N aqueous HClsolution. After the addition of brine, the organic layer was separatedand the aqueous layer was extracted with ethyl acetate. The combinedorganic layers were dried over anhydrous sodium sulfate, concentratedunder reduced pressure and purified by silica gel column chromatography.The combined fractions were concentrated under reduced pressure andtriturated with cold water to provide the title compound as an off-whitesolid (0.63 g). LCMS m/z=178.2 [M+H]⁺; ¹H NMR (400 MHz, CDCl₃) δ ppm3.92 (s, 3H), 6.55 (bs, 1H), 7.04 (d, J=8.72 Hz, 1H), 8.15 (dd, J=8.72,2.15 Hz, 1H), 8.23 (d, J=1.89 Hz, 1H).

Step B: Preparation of Methyl3-Cyano-4-(trifluoromethylsulfonyloxy)benzoate

To a suspension of methyl 3-cyano-4-hydroxybenzoate (1.24 g, 7.0 mmol)in dichloromethane (35 mL) was added trifluoromethanesulfonic anhydride(1.8 mL, 10.7 mmol), diisopropylethylamine (1.8 mL, 10.3 mmol), andN,N-dimethylaminopyridine (0.21 g, 1.75 mmol) at 0° C. The mixture wasstirred at room temperature overnight. The reaction was quenched with 1N aqueous HCl solution. The organic layer was separated and the aqueouslayer was extracted with dichloromethane. The combined organic layerswere dried over anhydrous magnesium sulfate, concentrated under reducedpressure and purified by silica gel column chromatography to provide thetitle compound as a brown oil (1.44 g). ¹H NMR (400 MHz, CDCl₃) δ ppm3.99 (s, 3H), 7.59 (d, J=8.84 Hz, 1H), 8.37 (dd, J=8.84, 2.15 Hz, 1H),8.44 (d, J=2.02 Hz, 1H).

Step C: Preparation of Methyl 3-Cyano-4-cyclohexylbenzoate

To a solution of methyl 3-cyano-4-(trifluoromethylsulfonyloxy)benzoate(0.7 g, 2.26 mmol) in tetrahydrofuran (30 mL) was added 0.5 Mcyclohexylzinc(II) bromide solution in tetrahydrofuran (13.6 mL, 6.8mmol) and bis(tri-tert-butylphosphine)palladium (0.058 g, 0.113 mmol) atroom temperature. The mixture was heated under reflux for 2 h. Themixture was allowed to cool to room temperature, quenched with saturatedaqueous sodium bicarbonate solution and filtered through Celite®. Thefiltrate was extracted with ethyl acetate. The organic layer was driedover anhydrous sodium sulfate, concentrated under reduced pressure andpurified by silica gel column chromatography to provide the titlecompound as an oil (0.24 g). ¹H NMR (400 MHz, CDCl₃) δ ppm 1.24-1.33 (m,1H), 1.42-1.53 (m, 4H), 1.77-1.84 (m, 1H), 1.86-1.95 (m, 4H), 2.99-3.08(m, 1H), 3.93 (s, 3H), 7.45 (d, J=8.34 Hz, 1H), 8.17 (dd, J=8.15, 1.71Hz, 1H), 8.27 (d, J=1.52 Hz, 1H).

Step D: Preparation of 2-Cyclohexyl-5-(hydroxymethyl)benzonitrile

To a solution of methyl 3-cyano-4-cyclohexylbenzoate (299.0 mg, 1.229mmol) in 1,4-dioxane (30 mL) was added 2 M lithium borohydride solutionin tetrahydrofuran (1.23 mL, 2.46 mmol). The mixture was heated underreflux for 2.5 h. The mixture was cooled to 0° C. and quenched with 1 Naqueous HCl solution slowly to pH 5. After the addition of brinesolution, the mixture was extracted with ethyl acetate. The organiclayer was dried over anhydrous sodium sulfate, concentrated underreduced pressure and purified by silica gel column chromatography toprovide the title compound as a white solid (190.5 mg). LCMS m/z=216.5[M+H]⁺; ¹H NMR (400 MHz, CDCl₃) δ ppm 1.23-1.33 (m, 1H), 1.41-1.52 (m,4H), 1.76-1.83 (m, 1H), 1.84-1.92 (m, 4H), 2.90-3.05 (m, 1H), 4.70 (d,J=5.81 Hz, 2H), 7.36 (d, J=8.21 Hz, 1H), 7.53 (dd, J=8.27, 1.71 Hz, 1H),7.61 (d, J=1.39 Hz, 1H).

Step E: Preparation of 5-(Chloromethyl)-2-cyclohexylbenzonitrile

To 2-cyclohexyl-5-(hydroxymethyl)benzonitrile (190.5 mg, 0.885 mmol) wasadded thionyl chloride (5.0 mL, 68.1 mmol). The mixture was heated at50° C. for 2 h and then room temperature overnight. The mixture waspoured into an ice and stirred for 5 min. The mixture was extracted withdichloromethane. The organic layer was washed with saturated aqueoussodium bicarbonate solution, dried over anhydrous sodium sulfate andconcentrated under reduced pressure to provide the title compound as awhite solid (184.9 mg). LCMS m/z=234.1 [M+H]⁺; ¹H NMR (400 MHz, CDCl₃) δppm 1.23-1.31 (m, 1H), 1.40-1.52 (m, 4H), 1.76-1.82 (m, 1H), 1.83-1.92(m, 4H), 2.93-3.03 (m, 1H), 4.55 (s, 2H), 7.37 (d, J=8.08 Hz, 1H), 7.55(dd, J=8.02, 1.83 Hz, 1H), 7.62 (d, J=1.64 Hz, 1H).

Step F: Preparation of Ethyl2-(7-(3-Cyano-4-cyclohexylbenzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetate

To a solution of ethyl2-(7-hydroxy-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetate (166.0mg, 0.641 mmol) and 5-(chloromethyl)-2-cyclohexylbenzonitrile (147.0 mg,0.629 mmol) in N,N-dimethylformamide (15 mL) was added cesium carbonate(246.0 mg, 0.755 mmol). The mixture was stirred at room temperature for41 h. The mixture was diluted with ethyl acetate, filtered throughCelite®, concentrated under reduced pressure and purified by silica gelcolumn chromatography to provide the title compound as a yellow foam(185.3 mg). LCMS m/z=457.5 [M+H]⁺; ¹H NMR (400 MHz, CDCl₃) δ ppm1.18-1.28 (m, 1H), 1.30 (t, J=7.20 Hz, 3H), 1.41-1.52 (m, 4H), 1.79 (dd,J=12.82, 1.33 Hz, 1H), 1.85-1.93 (m, 4H), 2.06-2.15 (m, 1H), 2.50 (dd,J=16.80, 11.24 Hz, 1H), 2.68-2.86 (m, 4H), 2.94-3.04 (m, 1H), 3.49-3.61(m, 1H), 4.16-4.25 (m, 2H), 5.06 (s, 2H), 6.82 (dd, J=8.78, 2.46 Hz,1H), 6.97 (d, J=2.53 Hz, 1H), 7.21 (d, J=8.72 Hz, 1H), 7.37 (d, J=8.21Hz, 1H), 7.62 (dd, J=8.15, 1.71 Hz, 1H), 7.71 (d, J=1.52 Hz, 1H), 8.47(bs, 1H).

Step G: Preparation of2-(7-(3-Cyano-4-cyclohexylbenzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)aceticAcid

To a solution of ethyl2-(7-(3-cyano-4-cyclohexylbenzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetate(185.3 mg, 0.406 mmol) in 1,4-dioxane (5 mL) was added 1 M aqueous LiOHsolution (1.22 mL, 1.22 mmol). The mixture was stirred at roomtemperature for 5 h. The mixture was concentrated under reducedpressure. The residue was dissolved in ethyl acetate and then acidifiedwith 1 N aqueous HCl acid solution to pH 4. The organic layer wasseparated, dried over anhydrous sodium sulfate and concentrated underreduced pressure. The concentrate was triturated with dichloromethane toprovide the title compound as a pink solid (98.9 mg). LCMS m/z=429.6[M+H]⁺; ¹H NMR (400 MHz, DMSO-d₆) δ ppm 1.21-1.31 (m, 1H), 1.33-1.54 (m,4H), 1.68-1.74 (m, 1H), 1.74-1.88 (m, 4H), 2.08 (dd, J=4.93, 3.54 Hz,1H), 2.34 (dd, J=16.04, 8.97 Hz, 1H), 2.61-2.76 (m, 4H), 2.81-2.89 (m,1H), 3.45 (bs, 1H), 5.07 (s, 2H), 6.70 (dd, J=8.78, 2.46 Hz, 1H), 6.91(d, J=2.40 Hz, 1H), 7.19 (d, J=8.72 Hz, 1H), 7.52 (d, J=8.08 Hz, 1H),7.72 (dd, J=8.15, 1.71 Hz, 1H), 7.81 (d, J=1.52 Hz, 1H), 10.46 (s, 1H),12.18 (bs, 1H).

Example 1.14 Preparation of2-(7-(4-Propyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)aceticAcid (Compound 18) Step A: Preparation of2-(7-(4-Propyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetate

To a solution of ethyl2-(7-(4-chloro-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetate(213.8 mg, 0.473 mmol) in tetrahydrofuran (5 mL) was added 0.5 Mpropylzinc bromide solution in tetrahydrofuran (4.7 mL, 2.4 mmol) andbis(tri-tert-butylphosphine)palladium (24.7 mg, 0.047 mmol). The mixturewas heated at 90° C. for 64 h. The mixture was allowed to cool to roomtemperature, quenched with saturated aqueous sodium bicarbonate solutionand filtered. The filtrate was extracted with ethyl acetate. The organiclayer was dried over anhydrous sodium sulfate, concentrated underreduced pressure and purified by silica gel column chromatography toprovide the title compound as a yellow foam (69.1 mg). LCMS m/z=460.5[M+H]⁺.

Step B: Preparation of2-(7-(3-Cyano-4-cyclohexylbenzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)aceticAcid

To a solution of ethyl2-(7-(4-propyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetate(75.8 mg, 0.165 mmol) in 1,4-dioxane (2 mL) was added 1 M aqueous LiOHsolution (0.495 mL, 0.495 mmol). The mixture was stirred at roomtemperature for 5 h. The mixture was then quenched with 1 N aqueous HClsolution to pH 5. After the addition of brine solution, the mixture wasextracted with ethyl acetate, dried over anhydrous sodium sulfate,concentrated under reduced pressure and purified by silica gel columnchromatography to provide the title compound as a purple foam (9.7 mg).LCMS m/z=432.5 [M+H]⁺; ¹H NMR (400 MHz, CD₃CN) δ ppm 0.98 (t, J=7.33 Hz,3H), 1.57-1.71 (m, 2H), 2.04-2.18 (m, 1H), 2.60 (d, J=7.45 Hz, 2H),2.66-2.83 (m, 5H), 3.46-3.57 (m, 1H), 5.10 (s, 2H), 6.76 (dd, J=8.78,2.46 Hz, 1H), 6.96 (d, J=2.40 Hz, 1H), 7.23 (d, J=8.84 Hz, 1H), 7.44 (d,J=7.96 Hz, 1H), 7.62 (d, J=7.96 Hz, 1H), 7.73 (s, 1H), 8.86 (bs, 1H).

Example 1.15 Preparation of2-(7-(4-Cyclobutyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)aceticAcid (Compound 21)

To a solution of ethyl2-(7-(4-chloro-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetate(202.8 mg, 0.449 mmol) in tetrahydrofuran (1 mL) was added 0.5 Mcyclobutylzinc(II) bromide solution in tetrahydrofuran (8.98 mL, 4.49mmol) and bis(tri-t-butylphosphine)palladium (46.8 mg, 0.090 mmol) atroom temperature. The mixture was heated at 90° C. for 63 h. The mixturewas then quenched with 1 N aqueous HCl solution and filtered throughCelite®. The filtrate was extracted with ethyl acetate. The organiclayer was washed with brine solution to remove excess HCl andconcentrated under reduced pressure. The residue was purified by HPLC.The combined fractions were triturated with saturated aqueous sodiumbicarbonate solution to basic solution and concentrated under reducedpressure. The residue was dissolved in ethyl acetate and acidified to pH4. The organic layer was washed with water until the aqueous layer wasneutral. The organic layer was dried over anhydrous sodium sulfate andconcentrated under reduced pressure to provide the title compound as apink solid (32.3 mg). LCMS m/z=444.6 [M+H]⁺; ¹H NMR (400 MHz, DMSO-d₆) δppm 1.78-1.87 (m, 1H), 1.93-2.02 (m, 1H), 2.03-2.12 (m, 1H), 2.16-2.28(m, 4H), 2.34 (dd, J=15.98, 9.03 Hz, 1H), 2.60-2.75 (m, 4H), 3.41-3.51(m, 1H), 3.74-3.84 (m, 1H), 5.12 (s, 2H), 6.70 (dd, J=8.78, 2.46 Hz,1H), 6.92 (d, J=2.40 Hz, 1H), 7.19 (d, J=8.84 Hz, 1H), 7.66-7.78 (m,3H), 10.46 (s, 1H), 12.20 (bs, 1H).

Example 1.16 Preparation of2-(7-(4-Cyclopropyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)aceticAcid (Compound 22)

To a solution of ethyl2-(7-(4-chloro-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetate(200.5 mg, 0444 mmol) in tetrahydrofuran (1 mL) was added 0.5 Mcyclopropylzinc(II) bromide solution in tetrahydrofuran (8.87 mL, 4.44mmol) and bis(tri-t-butylphosphine)palladium (46.3 mg, 0.089 mmol) atroom temperature. The mixture was heated at 90° C. for 63 h. The mixturewas then quenched with 1 N aqueous HCl solution and filtered throughCelite®. The filtrate was extracted with ethyl acetate. The organiclayer was washed with brine solution to remove excess HCl andconcentrated under reduced pressure. The residue was purified by HPLC.The combined fractions were triturated with saturated aqueous sodiumbicarbonate solution to basic solution and concentrated under reducedpressure. The residue was dissolved in ethyl acetate and acidified to pH4. The organic layer was washed with water until the aqueous layer wasneutral. The organic layer was dried over anhydrous sodium sulfate andconcentrated under reduced pressure to provide the title compound as alight brown solid (36.6 mg). LCMS m/z=430.5 [M+H]⁺; ¹H NMR (400 MHz,DMSO-d₆) δ ppm 0.74-0.88 (m, 2H), 0.98-1.07 (m, 2H), 2.02-2.12 (m, 2H),2.34 (dd, J=15.98, 9.03 Hz, 1H), 2.59-2.76 (m, 4H), 3.39-3.52 (m, 1H),5.10 (s, 2H), 6.69 (dd, J=8.72, 2.53 Hz, 1H), 6.91 (d, J=2.40 Hz, 1H),7.10-7.26 (m, 2H), 7.61 (d, J=8.34 Hz, 1H), 7.72 (d, J=1.14 Hz, 1H),10.45 (s, 1H), 12.20 (bs, 1H).

Example 1.17 Preparation of2-(7-((6-Cyclopentyl-5-(trifluoromethyl)pyridin-3-yl)methoxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)aceticAcid (Compound 19) Step A: Preparation of2-Chloro-5-(chloromethyl)-3-(trifluoromethyl)pyridine

To a cold solution of5-(chloromethyl)-2-methoxy-3-(trifluoromethyl)pyridine (0.3 g, 1.33mmol) in DMF (0.6 mL) was added dropwise POCl₃ (1.02 g, 6.65 mmol). Thereaction was stirred for 1 h at 100° C. in a sealed tube. The reactionwas cooled to room temperature, and poured onto ice water (10 mL). Thereaction was extracted with DCM (thrice) and the combined organic layerwas washed with water and brine, dried with MgSO₄ and concentrated. Theresidue was purified by silica gel column chromatography to provide thetitle compound (0.20 g). LCMS m/z=230.1 [M+H]⁺. ¹H NMR (400 MHz, CDCl₃)δ ppm 4.62 (s, 2H), 8.06 (d, J=2.3 Hz, 1H), 8.57 (d, J=2.3 Hz, 1H).

Step B: Preparation of Ethyl2-[7-{(6-Chloro-5-trifluoromethyl)pyridine-3-yl}methoxy-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl]acetate

From ethyl2-(7-hydroxy-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetate and2-chloro-5-(chloromethyl)-3-(trifluoromethyl)pyridine, in a similarmanner to the one described in Example 1.4, Step E, the title compoundwas obtained. LCMS m/z=453.4 [M+H]⁺. ¹H NMR (400 MHz, CDCl₃) δ ppm 1.29(t, J=7.1 Hz, 3H), 2.06-2.14 (m, 1H), 2.50 (dd, J=16.9, 11.2 Hz, 1H),2.71-2.86 (m, 4H), 3.50-3.58 (m, 1H), 4.16-4.24 (m, 2H), 5.14 (s, 2H),6.81 (dd, J=8.7 and 2.4 Hz, 1H), 6.97 (d, J=2.4 Hz, 1H), 7.22 (d, J=8.6Hz, 1H), 8.14 (d, J=2.1 Hz, 1H), 8.52 (bs, 1H), 8.63 (d, J=2.1 Hz, 1H).

Step C: Preparation of ethyl2-[7-{(6-Cyclopentyl-5-trifluoromethyl)pyridine-3-yl}methoxy-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl]acetate

From ethyl2-[7-{(6-chloro-5-trifluoromethyl)pyridine-3-yl}methoxy-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl]acetateand cyclopentylzinc(II) bromide, in a similar manner to the onedescribed in Example 1.8, Step C, the title compound was obtained. LCMSm/z=487.4 [M+H]⁺.

Step D: Preparation of2-[7-{(6-Cyclopentyl-5-trifluoromethyl)pyridine-3-yl}methoxy-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl]aceticAcid

The title compound was obtained from ethyl2-[7-{(6-cyclopentyl-5-trifluoromethyl)pyridine-3-yl}methoxy-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl]acetate,in a similar manner to the one described in Example 1.4 Step F. LCMSm/z=459.5 [M+H]⁺. ¹H NMR (400 MHz, CDCl₃) δ ppm 1.68-1.75 (m, 2H),1.87-1.95 (m, 4H), 1.98-2.05 (m, 2H), 2.10-2.19 (m, 1H), 2.62 (dd,J=17.1 and 10.8 Hz, 1H), 2.72-2.88 (m, 4H), 3.44-3.50 (m, 1H), 3.55-3.62(m, 1H), 5.11 (s, 2H), 6.83 (dd, J=8.8 and 2.4 Hz, 1H), 7.01 (d, J=2.4Hz, 1H), 7.22 (d, J=8.7 Hz, 1H), 8.00 (d, J=1.8 Hz, 1H), 8.35 (bs, 1H),8.80 (d, J=1.8 Hz, 1H).

Example 1.18 Preparation of2-(7-((6-(Pyrrolidin-1-yl)-5-(trifluoromethyl)pyridin-3-yl)methoxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)aceticAcid (Compound 23) Step A: Preparation of ethyl2-[7-{(6-Pyrrolidin-1-yl)-5-(trifluoromethyl)pyridine-3-yl}methoxy-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl]acetate

Ethyl2-[7-{(6-chloro-5-trifluoromethyl)pyridine-3-yl}methoxy-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl]acetate(60 mg, 0.132 mmol), pyrrolidine (47 mg, 0.66 mmol), Et₃N (67 mg, 0.66mmol), and IPA (0.7 mL) in a heavy welled tube was heated undermicrowave irradiation at 180° C. for 2 h. The mixture was purified byHPLC to provide the title compound (20 mg). LCMS m/z=488.5 [M+H]⁺.

Step B: Preparation of2-[7-{(6-Pyrrolidin-1-yl)-5-(trifluoromethyl)pyridine-3-yl}methoxy-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl]aceticAcid

From ethyl2-[7-{(6-pyrrolidin-1-yl)-5-(trifluoromethyl)pyridine-3-yl}methoxy-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl]acetate,in a similar manner to the one described in Example 1.4, Step F, thetitle compound was obtained. LCMS m/z=460.6 [M+H]⁺. ¹H NMR (400 MHz,CDCl₃) δ ppm 1.92-1.96 (m, 4H), 2.10-2.17 (m, 1H), 2.60 (dd, J=17.1,10.6 Hz, 1H), 2.74-2.87 (m, 4H), 3.55-3.62 (m, 5H), 4.97 (s, 2H), 6.80(dd, J=8.7, 2.4 Hz, 1H), 7.00 (d, J=2.4 Hz, 1H), 7.19 (d, J=8.6 Hz, 1H),7.91 (d, J=2.2 Hz, 1H), 8.30 (bs, 1H), 8.34 (d, J=2.2 Hz, 1H).

Example 1.19 Preparation of2-(7-((6-(3,3-Difluoropyrrolidin-1-yl)-5-(trifluoromethyl)pyridin-3-yl)methoxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)aceticAcid (Compound 20) Step A: Preparation of Ethyl2-(7-((6-(3,3-Difluoropyrrolidin-1-yl)-5-(trifluoromethyl)pyridin-3-yl)methoxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetate

From ethyl2-[7-{(6-chloro-5-trifluoromethyl)pyridine-3-yl}methoxy-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl]acetateand 3,3-difluoropyrrolidine hydrochloride, in a similar manner to theone described in Example 1.18, Step A, the title compound was obtained.LCMS m/z=524.4 [M+H]⁺.

Step B: Preparation of2-[7-{(6-(3,3-Difluoropyrrolidin-1-yl)-5-(trifluoromethyl)pyridine-3-yl}methoxy-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl]aceticAcid

Ethyl2-[7-{(6-(3,3-difluoropyrrolidin-1-yl)-5-(trifluoromethyl)pyridine-3-yl}methoxy-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl]acetatewas hydrolysized with 1 N LiOH in a similar manner to the one describedin Example 1.4, Step F to provide the title compound. LCMS m/z=496.4[M+H]⁺. ¹H NMR (400 MHz, CDCl₃) δ ppm 2.09-2.18 (m, 1H), 2.35-2.46 (m,2H), 2.60 (dd, J=17.1, 10.8 Hz, 1H), 2.74-2.88 (m, 4H), 3.54-3.61 (m,1H), 3.84 (t, J=7.3 Hz, 2H), 3.94 (t, J=13.4 Hz, 2H), 5.00 (s, 2H), 6.80(dd, J=8.6, 2.4 Hz, 1H), 6.99 (d, J=2.5 Hz, 1H), 7.20 (d, J=8.7 Hz, 1H),7.96 (d, J=2.2 Hz, 1H), 8.32 (bs, 1H), 8.39 (d, J=2.2 Hz, 1H).

Example 1.20 Preparation of2-(7-(4-(Methylsulfonyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)aceticacid (Compound 30) Step A: Preparation of Ethyl2-(7-(4-(Methylsulfonyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetate

To a stirred mixture of ethyl2-(7-hydroxy-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetate (50 mg,0.19 mmol) and cesium carbonate (94 mg, 0.29 mmol) in DMF (1.5 mL) wasadded 1-(bromomethyl)-4-(methylsulfonyl)benzene (72 mg, 0.29 mmol). Thereaction mixture was stirred at room temperature for 1 h, the solid wasfiltered. The filtrate was concentrated, and the residue was purified bypreparative TLC to give the title compound (40 mg) as an off-whitesolid. LCMS m/z=428.3 [M+H]⁺. ¹H NMR (400 MHz, CDCl₃) δ 1.30 (t, J=7.2Hz, 3H), 2.07-2.15 (m, 1H), 2.50 (dd, J=16.7 and 11.2 Hz, 1H), 2.70-2.86(m, 4H), 3.05 (s, 3H), 3.52-3.58 (m, 1H), 4.17-4.25 (m, 2H), 5.20 (s,2H), 6.84 (dd, J=8.8 and 2.4 Hz, 1H), 6.96 (d, J=2.1 Hz, 1H), 7.21 (d,J=8.8 Hz, 1H), 7.68 (d, J=8.1 Hz, 2H), 7.95 (d, J=8.3 Hz, 2H), 8.49 (s,1H).

Step B: Preparation of2-(7-(4-(Methylsulfonyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)aceticAcid

To the stirred solution of ethyl2-(7-(4-(methylsulfonyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetate(40 mg, 0.094 mmol) in dioxane was added 1M LiOH aqueous solution (0.47mL, 0.47 mmol). The reaction mixture was stirred at room temperature for24 h. The solvent was partly removed, diluted with water, and acidifiedwith HCl solution. The pinkish solid was collected and dried to give thetitle compound (26 mg). LCMS m/z=400.4 [M+H]⁺. ¹H NMR (400 MHz, DMSO-d₆)δ 2.04-2.08 (m, 1H), 2.35 (dd, J=16.0 and 9.0 Hz, 1H), 2.63-2.75 (m,4H), 3.20 (s, 3H), 3.45-3.50 (m, 1H), 5.21 (s, 2H), 6.73 (dd, J=8.7 and2.4 Hz, 1H), 6.92 (d, J=2.4 Hz, 1H), 7.20 (d, J=8.7 Hz, 1H), 7.71 (d,J=8.3 Hz, 2H), 7.92 (d, J=8.3 Hz, 2H), 10.47 (s, 1H), 12.18 (s, 1H).

Example 1.21 Preparation of2-(7-(4-(Cyclohexylmethyl)-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)aceticacid (Compound 28) Step A: Preparation of Methyl4-(Cyclohexylmethyl)-3-(trifluoromethyl)benzoate

To a stirred solution of methyl 4-chloro-3-(trifluoromethyl)benzoate(238 mg, 1.0 mmol) and bis(tri-t-butylphosphine)palladium (0) (51 mg,0.10 mmol) in THF (2 mL) was added (cyclohexylmethyl)zinc(II) bromide (6mL, 3.00 mmol) at room temperature. The reaction mixture was heated atreflux for 2 h, quenched with saturated NaHCO₃ solution, filteredthrough Celite. The filtrate was extracted with ethyl acetate. Thecombined organics were dried and concentrated, and the residue waspurified by column chromatography to give the title compound (280 mg) ascolorless oil. LCMS m/z=301.4 [M+H]⁺. ¹H NMR (400 MHz, CDCl₃) δ0.96-1.06 (m, 2H), 1.14-1.22 (m, 3H), 1.62-1.72 (m, 6H), 2.71 (d, J=6.7Hz, 2H), 3.94 (s, 3H), 7.39 (d, J=8.1 Hz, 1H), 8.10 (dd, J=8.0 and 1.5Hz, 1H), 8.30 (d, J=1.4 Hz, 1H).

Step B: Preparation of(4-(Cyclohexylmethyl)-3-(trifluoromethyl)phenyl)methanol

To a stirred solution of methyl4-(cyclohexylmethyl)-3-(trifluoromethyl)benzoate (280 mg, 0.93 mmol) indioxane (8 mL) was added 2 M lithium borohydride in THF solution (0.93mL, 1.86 mmol). The reaction mixture was heated at 80° C. for 2 h,cooled down, poured into water, acidified with HCl solution to pH 4, andextracted with ethyl acetate. The combined organics were washed withsaturated NaHCO₃ solution and water, dried and concentrated. The residuewas purified by silica gel column chromatography to give the titlecompound (190 mg) as a colorless oil. ¹H NMR (400 MHz, CDCl₃) δ0.96-1.06 (m, 2H), 1.14-1.22 (m, 3H), 1.62-1.72 (m, 6H), 2.67 (d, J=6.7Hz, 2H), 4.71 (d, J=5.7 Hz, 2H), 7.29 (d, J=7.9 Hz, 1H), 7.45 (dd, J=8.0and 1.6 Hz, 1H), 7.62 (d, J=1.6 Hz, 1H).

Step C: Preparation of4-(Bromomethyl)-1-(cyclohexylmethyl)-2-(trifluoromethyl)benzene

To a stirred solution of(4-(cyclohexylmethyl)-3-(trifluoromethyl)phenyl)methanol (80 mg, 0.29mmol) in dry DCM (1 mL) was added tribromophosphine (11 μL, 0.12 mmol)at 0° C. The reaction mixture was slowly warmed to room temperature andstirred for 1 h, poured into water, extracted with DCM. The combinedorganics were washed with saturated NaHCO₃ solution and brine, dried andconcentrated. The residue was purified by column chromatography to givethe title compound (80 mg) as a colorless oil. ¹H NMR (400 MHz, CDCl₃) δ0.96-1.06 (m, 2H), 1.14-1.22 (m, 3H), 1.55-1.72 (m, 6H), 2.65 (d, J=6.5Hz, 2H), 4.49 (s, 2H), 7.28 (d, J=8.0 Hz, 1H), 7.47 (dd, J=8.0 and 1.7Hz, 1H), 7.63 (d, J=1.6 Hz, 1H).

Step D: Preparation of Ethyl2-(7-(4-(Cyclohexylmethyl)-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetate

To a stirred reaction mixture of ethyl2-(7-hydroxy-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetate (40 mg,0.15 mmol) and cesium carbonate (75 mg, 0.23 mmol) in DMF (1 mL) wasadded 4-(bromomethyl)-1-(cyclohexylmethyl)-2-(trifluoromethyl)benzene(78 mg, 0.23 mmol). The reaction mixture was stirred at room temperaturefor 1 h. The solid was filtered and washed with ethyl acetate. Thecombined filtrate was concentrated, and the residue was purified bypreparative TLC to give the title compound (40 mg) as an oil. LCMSm/z=514.5 [M+H]⁺. ¹H NMR (400 MHz, CDCl₃) δ 0.96-1.06 (m, 2H), 1.14-1.22(m, 3H), 1.30 (t, J=7.2 Hz, 3H), 1.55-1.72 (m, 6H), 2.07-2.15 (m, 1H),2.50 (dd, J=16.7 and 11.2 Hz, 1H), 2.66 (d, J=6.8 Hz, 2H), 2.70-2.86 (m,4H), 3.52-3.58 (m, 1H), 4.18-4.25 (m, 2H), 5.08 (s, 2H), 6.85 (dd, J=8.8and 2.4 Hz, 1H), 7.00 (d, J=2.4 Hz, 1H), 7.21 (d, J=8.8 Hz, 1H), 7.31(d, J=7.9 Hz and 1.6 Hz, 1H), 7.55 (d, J=7.8 Hz, 1H), 7.72 (d, J=1.6 Hz,1H), 8.46 (s, 1H).

Step E: Preparation of2-(7-(4-(Cyclohexylmethyl)-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)aceticAcid

To a stirred solution of ethyl2-(7-(4-(cyclohexylmethyl)-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetate(40 mg, 0.078 mmol) in dioxane was added 1 M LiOH aqueous solution (0.39mL, 0.39 mmol). The reaction mixture was stirred at room temperature for5 h. The solvent was partly removed, then diluted with water, acidifiedwith HCl solution. The pinkish solid was collect and dried to give thetitle compound (19.5 mg). LCMS m/z=486.3 [M+H]⁺. ¹H NMR (400 MHz,DMSO-d₆) δ 0.96-1.06 (m, 2H), 1.14-1.22 (m, 3H), 1.55-1.70 (m, 6H),2.05-2.10 (m, 1H), 2.40 (dd, J=16.0 and 9.0 Hz, 1H), 2.62-2.75 (m, 6H),3.42-3.50 (m, 1H), 5.12 (s, 2H), 6.72 (dd, J=8.7 and 2.4 Hz, 1H), 6.94(d, J=2.4 Hz, 1H), 7.21 (d, J=8.8 Hz, 1H), 7.45 (d, J=8.0 Hz, 1H), 7.66(d, J=7.8 Hz, 1H), 7.75 (s, 1H), 10.48 (s, 1H), 12.20 (br, 1H).

Example 1.22 Preparation of2-(7-(4-(Ethylamino)-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)aceticacid (Compound 27)

To a mixture of ethyl2-(7-(4-chloro-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetate(50 mg, 0.11 mmol) in dioxane was added2-dicyclohexylphosphino-2′-(N,N-dimethylamino)biphenyl (4.4 mg, 0.011mmol), Pd₂dba₃ (5 mg, 5.5 μmol, 2 M ethanamine in THF (0.28 mL, 0.55mmol) and sodium tert-butoxide (21 mg, 0.22 mmol). The reaction mixturewas heated at 120° C. for 2 h under microwave irradiation, quenched bysaturated NH₄Cl solution and extracted with ethyl acetate. The combinedorganics were dried and concentrated. The residue was purified first bypreparative TLC followed by preparative HPLC to give the title compound(7 mg) as a white solid. LCMS m/z=433.5 [M+H]⁺. ¹H NMR (400 MHz, CDCl₃)δ 1.30 (t, J=7.1 Hz, 3H), 2.10-2.17 (m, 1H), 2.62 (dd, J=17.0 and 10.9Hz, 1H), 2.75-2.86 (m, 4H), 3.23 (q, J=7.1 Hz, 2H), 3.54-3.62 (m, 1H),4.96 (s, 2H), 6.73 (d, J=8.6 Hz, 1H), 6.83 (dd, J=8.8 and 2.4 Hz, 1H),7.00 (d, J=2.4 Hz, 1H), 7.19 (d, J=8.8 Hz, 1H), 7.46 (dd, J=8.5 and 1.7Hz, 1H), 7.53 (d, J=1.7 Hz, 1H), 8.28 (s, 1H).

Example 1.23 Preparation of2-(7-(4-(Cyclopropylmethoxy)-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)aceticacid (Compound 26) Step A: Preparation of Cyclopropylmethyl4-(Cyclopropylmethoxy)-3-(trifluoromethyl)benzoate

To a solution of 4-hydroxy-3-(trifluoromethyl)benzoic acid (0.483 g,2.343 mmol) in DMF (10 mL) was added Cs₂CO₃ (2.29 g, 7.03 mmol),followed by (bromomethyl)cyclopropane (0.568 mL, 5.86 mmol). Thereaction was stirred at 80° C. for 16 h. The mixture was filtered. Thefiltrate was concentrated under vacuum and taken up in EtOAc. Theorganic solution was washed with water (thrice), dried over MgSO₄ andconcentrated. The residue was purified by silica gel columnchromatography to give the title compound as an oil (0.643 g). LCMSm/z=315.3 [M+H]⁺. ¹H NMR (400 MHz, CDCl₃) δ ppm 0.33-0.44 (m, 4H),0.59-0.69 (m, 4H), 1.21-1.34 (m, 2H), 4.01 (d, J=6.57 Hz, 2H), 4.15 (d,J=7.20 Hz, 2H), 6.99 (d, J=8.84 Hz, 1H), 8.18 (dd, J=8.72, 2.15 Hz, 1H),8.28 (d, J=2.02 Hz, 1H).

Step B: Preparation of 4-(Cyclopropylmethoxy)-3-(trifluoromethyl)benzoicAcid

To a solution of cyclopropylmethyl4-(cyclopropylmethoxy)-3-(trifluoromethyl)benzoate (0.642 g, 2.043 mmol)in THF:MeOH (1:1, 10 mL) was added LiOH (1M, aq) (12.26 mmol). Thereaction was stirred overnight, quenched with HCl (1M, aq) and extractedwith EtOAc (twice). The combined extracts were dried over MgSO₄ andconcentrated to give the title compound as a white solid (0.502 g). LCMSm/z=261.1 [M+H]⁺. ¹H NMR (400 MHz, CDCl₃) δ ppm 0.39-0.46 (m, 2H),0.62-0.71 (m, 2H), 1.23-1.38 (m, 1H), 4.03 (d, J=6.57 Hz, 2H), 7.02 (d,J=8.72 Hz, 1H), 8.23 (dd, J=8.72, 2.15 Hz, 1H), 8.34 (d, J=2.02 Hz, 1H).

Step C: Preparation of(4-(Cyclopropylmethoxy)-3-(trifluoromethyl)phenyl)methanol

To a solution of 4-(cyclopropylmethoxy)-3-(trifluoromethyl)benzoic acidin THF (7 mL) at 0° C. was slowly added BH₃DMS (2.0 M in THF) (1.592 mL,3.18 mmol). After stirring for 0.5 h at 0° C., the reaction was allowedto return to room temperature and stirred overnight. The reactionmixture was slowly added to a saturated solution of NaHCO₃ at 0° C. andextracted with EtOAc (thrice). The combined extracts were dried overMgSO₄ and purified by silica gel column chromatography to give the titlecompound as a solid (0.358 g). ¹H NMR (400 MHz, CDCl₃) δ ppm 0.34-0.43(m, 2H), 0.57-0.67 (m, 2H), 1.22-1.32 (m, 1H), 3.94 (d, J=6.57 Hz, 2H),4.66 (s, 2H), 6.96 (d, J=8.46 Hz, 1H), 7.46 (dd, J=8.46, 2.02 Hz, 1H),7.57 (s, 1H).

Step D: Preparation of4-(Chloromethyl)-1-(cyclopropylmethoxy)-2-(trifluoromethyl)benzene

To a solution of(4-(cyclopropylmethoxy)-3-(trifluoromethyl)phenyl)methanol (0.258 g,1.454 mmol) in toluene (4 mL) was added thionyl chloride (0.637 mL, 8.72mmol). The reaction was stirred at 75° C. for 1.5 h. The reactionmixture was poured into ice water and extracted with hexanes (twice).The combined extracts were washed with NaHCO₃ (thrice), dried over MgSO₄and concentrated to give the title compound as a white solid (0.275 g).¹H NMR (400 MHz, CDCl₃) δ ppm 0.36-0.43 (m, 2H), 0.59-0.67 (m, 2H),1.21-1.34 (m, 1H), 3.95 (d, J=6.44 Hz, 2H), 4.56 (s, 2H), 6.95 (d,J=8.59 Hz, 1H), 7.48 (dd, J=8.53, 2.34 Hz, 1H), 7.58 (d, J=2.15 Hz, 1H).

Step E: Preparation of Ethyl2-(7-(4-(Cyclopropylmethoxy)-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetate

To a solution of ethyl2-(7-hydroxy-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetate (0.069 g,0.264 mmol) in DMF (1 mL) was added Cs₂CO₃ (0.086 g, 0.264 mmol)followed by4-(chloromethyl)-1-(cyclopropylmethoxy)-2-(trifluoromethyl)benzene(0.070 g, 0.264 mmol). The reaction mixture was stirred for 16 h andfiltered. The filtrate was concentrated under vacuum and purified bysilica gel column chromatography to give the title compound as a lightyellow oil (0.023 g). LCMS m/z=488.2 [M+H]⁺. ¹H NMR (400 MHz, CDCl₃) δppm 0.35-0.44 (m, 2H), 0.58-0.67 (m, 2H), 1.24-1.34 (m, 4H), 2.04-2.16(m, 1H), 2.51 (dd, J=16.74, 11.05 Hz, 1H), 2.68-2.90 (m, 4H), 3.49-3.61(m, 1H), 3.94 (d, J=6.44 Hz, 2H), 4.15-4.27 (m, 2H), 5.03 (s, 2H), 6.82(dd, J=8.72, 2.40 Hz, 1H), 6.93-7.02 (m, 2H), 7.20 (d, J=8.84 Hz, 1H),7.56 (dd, J=8.53, 1.96 Hz, 1H), 7.67 (d, J=1.89 Hz, 1H), 8.45 (bs, 1H).

Step F: Preparation of2-(7-(4-(Cyclopropylmethoxy)-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)aceticacid

To a solution of ethyl2-(7-(4-(cyclopropylmethoxy)-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetate(22.9 mg, 0.047 mmol) in dioxane was added 1M LiOH (0.188 mL, 0.188mmol). The reaction mixture was stirred 3 h, taken up in EtOAc andwashed with 1 M HCl. The EtOAc extract was dried over MgSO₄ andconcentrated. The residue was purified by preparative HPLC/MS to givethe title compound as a solid. LCMS m/z=460.3 [M+H]⁺. ¹H NMR (400 MHz,DMSO-d₆) δ ppm 0.30-0.38 (m, 2H), 0.52-0.59 (m, 2H), 1.17-1.27 (m, 1H),2.03-2.13 (m, 1H), 2.35 (dd, J=15.98, 8.91 Hz, 1H), 2.59-2.76 (m, 4H),3.99 (d, J=6.69 Hz, 2H), 5.05 (s, 2H), 6.69 (dd, J=8.84, 2.40 Hz, 1H),6.91 (d, J=2.53 Hz, 1H), 7.19 (d, J=8.72 Hz, 1H), 7.24 (d, J=8.46 Hz,1H), 7.62-7.71 (m, 2H), 10.45 (s, 1H).

Example 1.24 Preparation of2-(7-(4-(Cyclopentyloxy)-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)aceticacid (Compound 24) Step A: Preparation of Cyclopentyl4-(Cyclopentyloxy)-3-(trifluoromethyl)benzoate

From bromocyclopropane, the title compound was prepared using a similarmethod as described in Example 1.23, Step A to give an oil. LCMSm/z=343.4 [M+H]⁺. ¹H NMR (400 MHz, CDCl₃) δ ppm 1.58-1.72 (m, 4H),1.75-1.88 (m, 6H), 1.88-2.02 (m, 6H), 4.87-4.99 (m, 1H), 5.32-5.45 (m,1H), 7.00 (d, J=8.72 Hz, 1H), 8.12 (dd, J=8.72, 2.02 Hz, 1H), 8.20 (d,J=1.89 Hz, 1H).

Step B: Preparation of 4-(Cyclopentyloxy)-3-(trifluoromethyl)benzoicAcid

From cyclopentyl 4-(cyclopentyloxy)-3-(trifluoromethyl)benzoate, thetitle compound was prepared using a similar method as described inExample 1.23, Step B to give a white solid. LCMS m/z=275.4 [M+H]⁺.

Step C: Preparation of(4-(Cyclopentyloxy)-3-(trifluoromethyl)phenyl)methanol

From 4-(cyclopentyloxy)-3-(trifluoromethyl)benzoic acid, the titlecompound was prepared using a similar method as described in Example1.23, Step C to give an oil. ¹H NMR (400 MHz, CDCl₃) δ ppm 1.61-1.68 (m,2H), 1.76-1.86 (m, 2H), 1.86-1.96 (m, 4H), 4.64 (s, 2H), 4.84-4.91 (m,1H), 6.98 (d, J=8.59 Hz, 1H), 7.45 (dd, J=8.59, 2.15 Hz, 1H), 7.55 (d,J=1.89 Hz, 1H).

Step D: Preparation of4-(Chloromethyl)-1-(cyclopentyloxy)-2-(trifluoromethyl)benzene

From (4-(cyclopentyloxy)-3-(trifluoromethyl)phenyl)methanol, the titlecompound was prepared using a similar method as described in Example1.23, Step D to give an oil. ¹H NMR (400 MHz, CDCl₃) δ ppm 1.58-1.70 (m,2H), 1.76-1.86 (m, 2H), 1.86-1.95 (m, 4H), 4.56 (s, 2H), 4.84-4.90 (m,1H), 6.96 (d, J=8.59 Hz, 1H), 7.47 (dd, J=8.53, 2.34 Hz, 1H), 7.57 (d,J=2.15 Hz, 1H).

Step E: Preparation of Ethyl2-(7-(4-(Cyclopentyloxy)-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetate

From 4-(chloromethyl)-1-(cyclopentyloxy)-2-(trifluoromethyl)benzene, thetitle compound was prepared using a similar method as described inExample 1.23, Step E to give an oil. LCMS m/z=502.4 [M+H]⁺. ¹H NMR (400MHz, CDCl₃) δ ppm 1.30 (t, J=7.14 Hz, 3H), 1.57-1.69 (m, 2H), 1.76-1.97(m, 6H), 2.05-2.16 (m, 1H), 2.51 (dd, J=16.74, 11.18 Hz, 1H), 2.68-2.89(m, 4H), 3.49-3.61 (m, 1H), 4.14-4.28 (m, 2H), 4.84-4.91 (m, 1H), 5.02(s, 2H), 6.83 (dd, J=8.78, 2.46 Hz, 1H), 6.94-7.04 (m, 2H), 7.21 (d,J=8.72 Hz, 1H), 7.55 (dd, J=8.59, 2.02 Hz, 1H), 7.65 (d, J=2.02 Hz, 1H),8.45 (bs, 1H).

Step F: Preparation of2-(7-(4-(Cyclopentyloxy)-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)aceticacid

The title compound was prepared using a similar method as described inExample 1.23, Step F to give a solid. LCMS m/z=474.4 [M+H]⁺. ¹H NMR (400MHz, DMSO-d₆) δ ppm 1.53-1.79 (m, 6H), 1.82-1.97 (m, 2H), 2.00-2.15 (m,1H), 2.35 (dd, J=15.92, 8.97 Hz, 1H), 2.58-2.79 (m, 4H), 3.41-3.51 (m,1H), 4.94-5.08 (m, 3H), 6.69 (dd, J=8.78, 2.46 Hz, 1H), 6.92 (d, J=2.40Hz, 1H), 7.19 (d, J=8.72 Hz, 1H), 7.25 (d, J=9.22 Hz, 1H), 7.61-7.72 (m,2H), 10.45 (s, 1H), 12.18 (bs, 1H)

Example 1.25 Preparation of2-(7-(4-Cyano-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)aceticacid (Compound 25) Step A: Preparation of4-(Hydroxymethyl)-2-(trifluoromethyl)benzonitrile

To a solution of (4-chloro-3-(trifluoromethyl)phenyl)methanol (0.300 g,1.425 mmol) in DMA was added dicyanozinc (0.335 g, 2.85 mmol) andtetrakis(triphenylphosphine) palladium (0) (0.165 g, 0.142 mmol). Thereaction flask was degassed and charged with nitrogen, then heated at150° C. for 6 h under microwave irradiation. The reaction mixture waspoured into water and extracted with EtOAc. The EtOAc extract was washedwith brine, dried over MgSO₄ and purified by silica gel columnchromatography to give the title compound as a white solid (0.105 g).LCMS m/z=202.1 [M+H]⁺. ¹H NMR (400 MHz, CDCl₃) δ ppm 1.98 (bs, 1H), 4.87(s, 2H), 7.69 (d, J=0.88 Hz, 1H), 7.77-7.88 (m, 2H)

Step B: Preparation of 4-(chloromethyl)-2-(trifluoromethyl)benzonitrile

From 4-(hydroxymethyl)-2-(trifluoromethyl)benzonitrile, the titlecompound was prepared using a similar method as described in Example1.23, Step D to give an oil. LCMS m/z=220.2 [M+H]⁺. ¹H NMR (400 MHz,DMSO-d₆) δ ppm 4.65 (s, 2H), 7.73 (d, J=1.39 Hz, 1H), 7.84 (d, J=7.71Hz, 2H).

Step C: Preparation of Ethyl2-(7-(4-Cyano-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetate

From 4-(chloromethyl)-2-(trifluoromethyl)benzonitrile, the titlecompound was prepared using a similar method as described in Example1.23 Step E to give an oil. LCMS m/z=443.3 [M+H]⁺. ¹H NMR (400 MHz,CDCl₃) δ ppm 1.30 (t, J=7.14 Hz, 3H), 2.03-2.18 (m, 1H), 2.50 (dd,J=16.80, 11.24 Hz, 1H), 2.68-2.91 (m, 4H), 3.48-3.63 (m, 1H), 4.13-4.28(m, 2H), 5.21 (s, 2H), 6.83 (dd, J=8.72, 2.53 Hz, 1H), 6.96 (d, J=2.40Hz, 1H), 7.23 (d, J=8.72 Hz, 1H), 7.73-7.81 (m, 1H), 7.81-7.88 (m, 1H),7.91 (s, 1H), 8.52 (bs, 1H).

Step D: Preparation of2-(7-(4-cyano-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)aceticacid

The title compound was prepared using a similar method as described inExample 1.23, Step F to give a solid. LCMS m/z=415.4 [M+H]⁺. ¹H NMR (400MHz, DMSO-d₆) δ ppm 2.01-2.14 (m, 1H), 2.35 (dd, J=15.98, 9.03 Hz, 1H),2.57-2.78 (m, 4H), 3.38-3.53 (m, 1H), 5.29 (s, 2H), 6.75 (dd, J=8.78,2.46 Hz, 1H), 6.94 (d, J=2.53 Hz, 1H), 7.22 (d, J=8.72 Hz, 1H), 7.97 (s,1H), 8.07 (s, 1H), 8.19 (d, J=7.96 Hz, 1H), 10.50 (s, 1H), 12.18 (bs,1H)

Example 1.26 Preparation of2-(7-(4-Carbamoyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)aceticacid (Compound 29)

To a solution of2-(7-(4-cyano-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)aceticacid (9.0 mg, 0.022 mmol) in dioxane (1 mL) was added 1M LiOH (aq) (3.0mL). The reaction was stirred at 50° C. for 48 h. 1M HCl (aq) was addedto adjust pH to 3. The mixture was extracted with EtOAc. The EtOAcextract was dried over MgSO₄ and the residue was purified by preparativeHPLC/MS to give the title compound as a solid (2.1 mg). LCMS m/z=433.2[M+H]⁺. ¹H NMR (400 MHz, DMSO-d₆) δ ppm 2.01-2.14 (m, 1H), 2.32-2.42 (m,1H), 2.57-2.78 (m, 4H), 5.20 (s, 2H), 6.72 (d, J=11.37 Hz, 1H), 6.93 (d,J=2.40 Hz, 1H), 7.20 (d, J=8.72 Hz, 1H), 7.50-7.59 (m, 2H), 7.76 (d,J=7.58 Hz, 1H), 7.82 (s, 1H), 7.91 (s, 1H), 10.47 (s, 1H), 12.17 (bs,1H).

Example 1.27 Preparation of2-(7-(4-(Pyrazin-2-yl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)aceticacid (Compound 31) Step A: Preparation of(4-(Pyrazin-2-yl)phenyl)methanol

A mixture of 2-chloropyrazine (0.230 ml, 2.62 mmol),4-(hydroxymethyl)phenylboronic acid (517 mg, 3.41 mmol),tetrakis(triphenylphosphine)palladium (0) (303 mg, 0.262 mmol) and 2 Mpotassium phosphate aqueous solution (2.62 ml, 5.24 mmol) in dioxane (10mL) was heated at 80° C. overnight under nitrogen. The mixture wascooled down, poured into water and extracted with ethyl acetate. Thecombined organic layers were dried and concentrated. The residue waspurified by preparative HPLC to give the title compound (350 mg) as anoff-white solid. LCMS m/z=187.0 [M+H]⁺. ¹H NMR (400 MHz, CDCl₃) δ 4.79(s, 2H), 7.52 (d, J=8.1 Hz, 2H), 8.02 (d, J=8.1 Hz, 2H), 8.51 (d, J=2.5Hz, 1H), 8.63 (dd, J=2.4 and 1.6 Hz, 1H), 9.03 (d, J=1.6 Hz, 1H).

Step B: Preparation of 4-(Pyrazin-2-yl)benzyl methanesulfonate

To a stirred solution of (4-(pyrazin-2-yl)phenyl)methanol (40 mg, 0.22mmol) and DIEA (56 μL, 0.32 mmol) in DCM (1 mL) was addedmethanesulfonyl chloride (29.5 mg, 0.258 mmol) at 0° C. The reactionmixture was stirred at that temperature for 1 h, poured into water, andextracted with DCM. The combined organics were dried and concentrated togive the title compound (50 mg) without further purification. LCMSm/z=265.1 [M+H]⁺.

Step C: Preparation of Ethyl2-(7-(4-(Pyrazin-2-yl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetate

To a mixture of ethyl2-(7-hydroxy-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetate (20 mg,0.077 mmol) and cesium carbonate (38 mg, 0.12 mmol) in DMF (1 mL) wasadded 4-(pyrazin-2-yl)benzyl methanesulfonate (41 mg, 0.15 mmol). Thereaction mixture was stirred at room temperature overnight. The solidwas filtered, and the filtrate was concentrated. The residue waspurified by preparative TLC to give the title compound (15 mg). LCMSm/z=428.3 [M+H]⁺. ¹H NMR (400 MHz, CDCl₃) δ 1.30 (t, J=7.1 Hz, 3H),2.07-2.14 (m, 1H), 2.50 (dd, J=16.7 and 11.2 Hz, 1H), 2.70-2.87 (m, 4H),3.50-3.57 (m, 1H), 4.18-4.24 (m, 2H), 5.18 (s, 2H), 6.87 (dd, J=8.8 and2.4 Hz, 1H), 7.01 (d, J=2.3 Hz, 1H), 7.21 (d, J=8.7 Hz, 1H), 7.62 (d,J=8.2 Hz, 2H), 8.03 (d, J=8.3 Hz, 2H), 8.46 (s, 1H), 8.51 (d, J=2.5 Hz,1H), 8.64 (dd, J=2.3 and 1.6 Hz, 1H), 9.04 (d, J=1.4 Hz, 1H).

Step D: Preparation of2-(7-(4-(Pyrazin-2-yl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)aceticAcid

To a stirred solution of ethyl2-(7-(4-(pyrazin-2-yl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetate(15 mg, 0.035 mmol) in dioxane (1 mL) was added 1 M lithium hydroxidesolution (0.175 mL, 0.175 mmol). The reaction mixture was stirred atroom temperature for 5 h and acidified with HCl solution. The mixturewas purified by HPLC to give the title compound (8 mg) as a pinkishsolid. LCMS m/z=400.4 [M+H]⁺. ¹H NMR (400 MHz, DMSO-d₆) δ 2.04-2.12 (m,1H), 2.35 (dd, J=16.0 and 9.0 Hz, 1H), 2.62-2.75 (m, 4H), 3.44-3.50 (m,1H), 5.17 (s, 2H), 6.74 (dd, J=8.7 and 2.4 Hz, 1H), 6.95 (d, J=2.4 Hz,1H), 7.21 (d, J=8.7 Hz, 1H), 7.62 (d, J=8.2 Hz, 2H), 8.15 (d, J=8.3 Hz,2H), 8.61 (d, J=2.5 Hz, 1H), 8.72 (dd, J=2.3 and 1.6 Hz, 1H), 9.26 (d,J=1.5 Hz, 1H), 10.47 (s, 1H), 12.20 (s, 1H).

Example 1.28 Preparation of2-(7-(4-(1,2,3-Thiadiazol-4-yl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)aceticacid (Compound 32) Step A: Preparation of Ethyl2-(7-(4-(1,2,3-Thiadiazol-4-yl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetate

In a 4 mL vial were placed ethyl2-(7-hydroxy-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetate (64.8 mg,0.250 mmol), cesium carbonate (81 mg, 0.250 mmol), and4-(4-(bromomethyl)phenyl)-1,2,3-thiadiazole (63.8 mg, 0.250 mmol). DMA(1 mL) was added and the reaction was stirred at room temperatureovernight. The solid was removed by filtration and rinsed with EtOAc.The filtrate was concentrated under reduced pressure and the residue waspurified by silica gel column chromatography to give the title compound(42.7 mg). LCMS ink=434.2 [M+H]⁺.

Step B: Preparation of2-(7-(4-(1,2,3-Thiadiazol-4-yl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)aceticAcid

To a solution of ethyl2-(7-(4-(1,2,3-thiadiazol-4-yl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetate(42.7 mg, 0.030 mmol) in dioxane was added 1M LiOH (0.394 mL, 0.394mmol). The reaction was stirred overnight. The reaction mixture wastaken up in EtOAc and washed with 1 M HCl. The EtOAc extract was driedover MgSO₄ and concentrated. The residue was purified by silica gelcolumn chromatography to give the title compound as a solid. LCMSm/z=406.4 [M+H]⁺. ¹H NMR (400 MHz, DMSO-d₆) δ ppm 2.03-2.12 (m, 1H),2.36 (dd, J=16.04, 8.97 Hz, 1H), 2.58-2.79 (m, 4H), 3.41-3.51 (m, 1H),5.16 (s, 2H), 6.74 (dd, J=8.78, 2.46 Hz, 1H), 6.95 (d, J=2.40 Hz, 1H),7.21 (d, J=8.72 Hz, 1H), 7.63 (d, J=8.21 Hz, 2H), 8.15 (d, J=8.21 Hz,2H), 9.61 (s, 1H), 10.46 (s, 1H), 12.19 (bs, 1H).

Example 1.29 Preparation of2-(7-(4-Cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)aceticAcid (Compound 12) Step A: Preparation of Methyl4-Chloro-3-(trifluoromethyl)benzoate

To a solution of 4-chloro-3-(trifluoromethyl)benzoic acid (200 g, 891mmol) in MeOH (600 mL, 14.8 mol), sulfuric acid (27 mL, 445 mmol) wasadded. The mixture was stirred at reflux for 6 h, allowed to cool andthe solvent evaporated under reduce pressure. The resulting liquidresidue (˜250 mL) was poured onto ice water whereby a white suspensionformed. The solid was filtered and washed with 0.05 N NaOH (3×200 mL)followed by H₂O (3×200 mL). The solid was dried under vacuum for 16 hfollowed by 4 h at 40° C. to give the title compound as an off-whitesolid (197.0 g). ¹H NMR (400 MHz, CDCl₃) δ ppm, 3.98 (s, 3H), 7.62 (d,J=8.4 Hz, 1H), 8.16 (dd, J=8.8 Hz, 2.0 Hz, 1H), 8.39 (d, J=2.0 Hz, 1H).

Step B: Preparation of Methyl 4-Cyclopentyl-3-(trifluoromethyl)benzoate

To a solution of 4-chloro-3-(trifluoromethyl)benzoate (196.7 g, 824mmol) in THF (100 mL), cyclopentylzinc(II) bromide (1979 mL, 989 mmol)was added dropwise at 7.8° C. The temperature at the end of the additionrose to 22° C. bis(Tri-t-butylphosphine)palladium (21.07 g, 41.2 mmol)was added to the dark brown solution at the same temperature, and theresulting mixture was stirred at 70° C. for 8 h. The mixture was addedto saturated aqueous NaHCO₃ (100 mL) at 0° C., stirred at the sametemperature for 30 min and then at 22° C. for 2 h. The resultingsuspension was filtered through Celite and the filtrate concentratedunder vacuum. The solids were washed with EtOAc (3×300 mL), the filtratewas combined with the previous concentrate and the combined organicswere washed with H₂O (2×600 mL), brine (2×500 mL), dried (Na₂SO₄),decanted and concentrated under reduced pressure to give the titlecompound as an orange oil (227 g) without further purification. LCMSm/z=273.4 [M+H]⁺; ¹H NMR (400 MHz, CDCl₃) δ ppm 1.71-1.60 (m, 2H),1.83-1.75 (m, 2H), 1.95-1.87 (m, 2H), 2.21-2.11 (m, 2H), 3.46 (quintet,J=8.8 Hz, 1H), 3.97 (s, 3H), 7.58 (d, J=8.4 Hz, 1H), 8.18 (dd, J=8.0 Hz,1.6 Hz, 1H), 8.31 (d, J=1.6 Hz, 1H).

Step C: Preparation of (4-Cyclopentyl-3-(trifluoromethyl)phenyl)methanol

To a solution of 4-cyclopentyl-3-(trifluoromethyl)benzoate (224 g, 823mmol) in 1,4-dioxane (600 mL), LiBH₄ (494 mL, 987 mmol, 2 M solution inTHF) was added dropwise at 22° C. The resulting suspension was stirredat 85.5° C. for 5.5 h. The dark brown solution was cooled to 0° C. andthe pH adjusted to 5 by slowly adding 6 N HCl (130 mL). The layers wereseparated and to the aqueous phase H₂O (250 mL) and NaCl (20 g) added.The combined aqueous were extracted with EtOAc (2×250 mL). The EtOAclayer was added to the previously separated organic phase and thecombined organics were concentrated under reduced pressure. Theresulting suspension was filtered through a pad of Celite/Na₂SO₄ and thesolids were washed with EtOAc (3×400 mL). The combined organics wererotary evaporated and the dark brown oily residue was subjected tochromatography on silica to give the title compound as colorless liquid(110 g). LCMS m/z=243.3 [M−H]⁺; ¹H NMR (400 MHz, CDCl₃) δ ppm 1.67-1.55(m, 2H), 1.82-1.69 (m, 2H), 1.95-1.83 (m, 2H), 2.19-2.04 (m, 2H), 3.39(quintet, J=8.0 Hz, 1H), 4.72 (s, 2H), 7.55-7.46 (m, 2H), 7.62 (s, 1H).

Step D: Preparation of4-(Chloromethyl)-1-cyclopentyl-2-(trifluoromethyl)benzene

To (4-cyclopentyl-3-(trifluoromethyl)phenyl)methanol (110 g, 113 mmol),thionyl chloride (329 mL, 4.50 mol) was added dropwise at such a rate asto maintain the internal temperature between 10-25° C. (cooled withice-water). The resulting mixture was stirred at 50° C. for 3.5 hfollowed by 6 h at 25° C. The mixture was concentrated under reducedpressure and the resulting oily residue poured into ice-water (450 mL)under vigorous stirring. The layers were separated and the aqueous phaseextracted with CH₂Cl₂ (3×400 mL). The combined organic layers werewashed with saturated NaHCO₃ (400 mL), brine (2×400 mL), dried (Na₂SO₄),filtered over fresh Na₂SO₄, and concentrated in vacuo to give the titlecompound as a pale yellow oil (113.3 g). ¹H NMR (400 MHz, CDCl₃) δ ppm1.67-1.57 (m, 2H), 1.81-1.71 (m, 2H), 1.94-1.84 (m, 2H), 2.16-2.07 (m,2H), 3.39 (quintet, J=8.6 Hz, 1H), 4.61 (s, 2H), 7.49 (d, J=8.4 Hz, 1H),7.54 (dd, J=8.4 Hz, 1.6 Hz, 1H), 7.63 (d, J=1.6 Hz, 1H).

Step E: Preparation of Ethyl2-(7-Methoxy-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetate

To a solution of 2-iodo-4-methoxyaniline (20.0 g, 80 mmol),ethyl-2-(2-oxocyclopentyl)acetate (20.5 g, 120 mmol, 1.5 eq) andtetraethyl orthosilicate (21.7 g, 104 mmol, 1.3 eq) in anhydrous DMF(100 mL), was added pyridine p-toluenesulfonate (0.807 g, 3.21 mmol,0.04 eq). The dark brown solution was stirred at 135° C. for 5 h underN₂, allowed to cool to 100° C. and then added DIPEA (31.1 g, 241 mmol, 3eq) followed by Pd(OAc)₂ (0.541 g, 2.41 mmol, 0.03 eq). The resultingmixture was stirred at 120° C. for 22 h under N₂/concentrated underreduced pressure. The residue was taken up in DCM, filtered through aplug of silica and the solvent was evaporated under reduced pressure.The residue was purified by silica gel column chromatography to give thetitle compound. LCMS m/z=274.4 [M+H]⁺.

Step F: Preparation of Ethyl2-(7-Hydroxy-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetate

DCM (305 mL) was transferred to a 1 L 3-necked round-bottomed flask andcooled to −11° C. (internal) (ice acetone bath). BBr₃ (72.0 mL, 761mmol) was added to the DCM with stirring. A solution of ethyl2-(7-methoxy-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetate (41.62 g,152 mmol) in DCM (145 mL) was added in drops maintaining the internaltemperature at between −5 to 0° C. After the addition the reaction wasstirred for 1 h below 0° C. The reaction mixture was slowly poured intomixture of ice (400 mL) and saturated K₂CO₃ (400 mL) and stirred well(pH maintained at 9-7). The organic layer was separated, washed withbrine (1×100 mL), dried over MgSO₄, filtered and concentrated underreduced pressure. The residual brown oil was purified by a pad of silicagel to give the title compound (8.03 g). LCMS m/z=260.2.

Step G: Preparation of Ethyl2-(7-(4-Cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetate

In a 2 L, 3-necked, round-bottomed flask under nitrogen atmosphere wereplaced ethyl2-(7-hydroxy-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetate (55.85 g,215 mmol), cesium carbonate (84.2 g, 258 mmol),4-(chloromethyl)-1-cyclopentyl-2-(trifluoromethyl)benzene (68 g, 259mmol) in DMA (670 mL). The mixture was stirred 15 minutes at roomtemperature and heated at 50° C. overnight. The mixture was cooled downto room temperature and filtered. The filtrate was concentrated undervacuum. The residue was added hexanes (400 mL) and heated to 40° C. togive a dark solution. The solution was cooled down to room temperatureover the weekend. The mixture was concentrated in vacuo and dried undervacuum to give the title compound (129.7 g). LCMS m/z=486.2.

Step H: Preparation of2-(7-(4-Cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)aceticAcid

In a 3 L, 3-necked, round-bottomed flask was placed ethyl2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetate(139.4 g, 287 mmol) in dioxane (1.8 L). The mixture was added 2N lithiumhydroxide (0.431 L, 861 mmol) and heated to 45-55° C. for 3 h. Themixture was concentrated in vacuo. The residue was added MTBE/water andacidified with concentrated HCl (until pH3) while keeping thetemperature under 20° C. with an ice bath. The aqueous layer wasseparated and extracted with MTBE. The combined organic layers werewashed several times with water until pH3 at the end of the washes.Acetonitrile and water were added to the MTBE solution and the mixturewas concentrated in vacuo to give the title compound (130 g) withoutfurther purification. LCMS m/z=458.4.

Resolution Via Chiral HPLC (Conducted by Chiral Technologies Inc)

Column: normal phase preparative ChiralCel® OJH®

Eluent: CO₂/MeOH (75-25%) Gradient: Isocratic

Flow: 400 mL/min

Detector: 254 nm

Retention Times: 1^(st) enantiomer: 9.1 min (appears to correspond tothe 2^(nd) enantiomer purified under chiral HPLC conditions described inExample 1.4); 2^(nd) enantiomer: 13.9 min (appears to correspond to the1^(st) enantiomer purified under chiral HPLC conditions described inExample 1.4).

After chiral resolution, the respective purified fractions wereconcentrated to dryness.

To a solution of the 1^(st) enantiomer of Compound 12 as described inExample 1.29 in acetonitrile and ethanol was added (S)-1-phenethylamine(1 equivalent). The mixture was heated briefly and allowed toconcentrate by slow evaporation. A precipitate was formed, filtered anddried. Single crystals of the 1^(st) enantiomer of Compound 12 (asdescribed in Example 1.29) were obtained and subjected to X-raycrystallography analysis. They were observed to be as depicted in FIG.21.

Example 1.30 Preparation of2-(7-(4-Cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)aceticAcid (Compound 12) Step A: Preparation of1-(2-(Trifluoromethyl)phenyl)cyclopentanol

A solution of 1-bromo-2-(trifluoromethyl)benzene (0.5 g, 2.222 mmol) inanhydrous THF (10 mL) was cooled to −78° C. (thy ice IPA bath) underargon atmosphere. BuLi (2.5 M in hexanes, 1.068 mL, 2.67 mmol) was addedin drops with efficient stirring. The reaction mixture was stirred at−78° C. for 40 min. A solution of cyclopentanone (0.243 g, 2.89 mmol) inanhydrous THF (1.5 mL) was added slowly (in drops) at −78° C. Thereaction mixture was stirred at −78° C. for 30 min, gradually brought toroom temperature, and stirred for 1 h. The reaction mixture was cooledby an ice bath, quenched with water, and acidified to pH 4-5 by additionof concentrated HCl. The solvent was removed under reduced pressure. Theresidue was dissolved in methylene chloride, washed with water (2times), dried over Na₂SO₄, filtered and concentrated under reducedpressure. The residue was purified by silica gel chromatography to givethe title compound as an oil (250 mg). LCMS m/z=213.1 [M−H₂O+H]⁺.

Step B: Preparation of 1-Cyclopentyl-2-(trifluoromethyl)benzene

To a solution of 1-(2-(trifluoromethyl)phenyl)cyclopentanol (5.1 g,22.15 mmol) in ethanol (32 mL) was added 10% Pd—C (500 mg; Degussa; wet)and the mixture was hydrogenated overnight with a hydrogen balloon. Thereaction mixture was filtered through celite. The filtrate was pouredinto ice-water (100 mL) and extracted with CH₂Cl₂ (2×70 mL). Thecombined CH₂Cl₂ layer was washed with water (1×75 mL), dried overNa₂SO₄, filtered and the solvent was removed under reduced pressure togive the title compound (4.3 g). ¹H NMR (400 MHz, CDCl₃) δ ppm 1.58-1.67(m, 4H), 1.81-1.90 (m, 2H), 2.06-2.15 (m, 2H), 3.32-3.43 (m, 1H),7.22-7.26 (m, 1H), 7.45-7.51 (m, 2H), 7.58 (d, J=8 Hz, 1H).

Step C: Preparation of 4-Bromo-1-cyclopentyl-2-(trifluoromethyl)benzene

To a solution of 1-cyclopentyl-2-(trifluoromethyl)benzene (0.5 g, 2.334mmol) in acetic acid (2.5 mL) was added bromine (1.202 mL, 23.34 mmol).The mixture was stirred well, added concentrated H₂SO₄ (2.5 mL), andstirred at 40° C. for 1.5 h. The reaction mixture was poured intoice-water and extracted with CH₂Cl₂. The CH₂Cl₂ layer was washed withwater, followed by a solution of sodium thiosulfate, then with water.The organic layer was dried over Na₂SO₄ and the solvent was removedunder reduced pressure. The residue was purified by silica gelchromatography to give the title compound (250 mg). ¹H NMR (400 MHz,DMSO-d₆) δ ppm 1.52-1.75 (m, 4H), 1.78-1.88 (m, 2H), 1.95-2.04 (m, 2H),3.16-3.26 (m, 1H), 7.57 (d, J=8.4 Hz, 1H), 7.76 (d, J=2 Hz, 1H), 7.81(dd, J=8.4 Hz, 2 Hz, 1H).

Step D: Preparation of 4-Cyclopentyl-3-(trifluoromethyl)benzaldehyde

In a 15 mL round-bottomed flask were placed4-Bromo-1-cyclopentyl-2-(trifluoromethyl)benzene (0.186 g, 0.635 mmol)and anhydrous THF (1.86 mL) under argon atmosphere. The solution wasstirred well and cooled to −78° C. (dry ice IPA bath). BuLi (2.5 M inhexanes, 0.281 mL, 0.703 mmol) was added in drops (slowly) and thereaction mixture was stirred at low temperature for 25 min. AnhydrousDMF (0.1 mL, 0.766 mmol) was added in drops at −78° C. (slowly). Themixture was stirred at −78° C. for 20 min then in room temperature for30 min. The reaction was quenched with water, acidified with 2M HCl andextracted with EtOAc. The EtOAc layer was washed with water, dried overNa₂SO₄, and concentrated in vacuo to give the title compound as an oil(60 mg). LCMS m/z=243.3 [M+H]⁺. ¹H NMR (400 MHz, DMSO-d₆) δ ppm 1.55-1.7(m, 4H), 1.79-1.94 (m, 2H), 1.95-2.09 (m, 2H), 3.29-3.37 (m, 1H), 7.86(d, J=8 Hz, 1H), 8.12 (d, J=8 Hz, 1H), 8.16 (d, J=1.2 Hz, 1H), 10.46 (s,1H).

Step E: Preparation of (4-Cyclopentyl-3-(trifluoromethyl)phenyl)methanol

To a solution of 4-cyclopentyl-3-(trifluoromethyl)benzaldehyde (0.25 g,1.032 mmol) in ethanol (2.5 mL) was added sodium borohydride (0.047 g,1.238 mmol) and the mixture was stirred in room temperature for 2 h. Themixture was quenched with water, acidified with 6N HCl, diluted withmore water and extracted with CH₂Cl₂. The CH₂Cl₂ layer was washed withwater, dried over Na₂SO₄, filtered and concentrated in vacuo to give thetitle compound (0.22 g). LCMS m/z=227.5 [M−H₂O+H]⁺. ¹H NMR (400 MHz,DMSO-d₆) δ ppm 1.54-1.72 (in, 4H), −1.77-1.89 (m, 2H), 1.93-2.05 (m,2H), 3.19-3.28 (m, 1H), 4.52 (d, J=6 Hz, 2H), 5.28 (t, J=5.6 Hz, 1H),7.52-7.6 (m, 3H).

Step F: Preparation of4-(Chloromethyl)-1-cyclopentyl-2-(trifluoromethyl)benzene

To (4-cyclopentyl-3-(trifluoromethyl)phenyl)methanol (110 g, 113 mmol)thionyl chloride (329 mL, 4.50 mol, 10 eq) was added dropwise at such arate as to maintain the internal temperature between 10-25° C. (cooledwith ice-water). The resulting mixture was stirred at 50° C. for 3.5 hfollowed by 6 h at 25° C. The mixture was concentrated under reducedpressure and the resulting oily residue poured into ice-water (450 mL)under vigorous stirring. The layers were separated and the aqueous phaseextracted with CH₂Cl₂ (3×400 mL). The combined organic layers werewashed with saturated NaHCO₃ (400 mL), brine (2×400 mL), dried (Na₂SO₄),filtered over fresh Na₂SO₄, and concentrated in vacuo to afford4-(chloromethyl)-1-cyclopentyl-2-(trifluoromethyl)benzene as a paleyellow oil (113.3 g, 96%). ¹H NMR (400 MHz, CDCl₃) δ ppm 1.67-1.57 (m,2H), 1.81-1.71 (m, 2H), 1.94-1.84 (m, 2H), 2.16-2.07 (m, 2H), 3.39(quintet, J=8.6 Hz, 1H), 4.61 (s, 2H), 7.49 (d, J=8.4 Hz, 1H), 7.54 (dd,J=8.4 Hz, 1.6 Hz, 1H), 7.63 (d, J=1.6 Hz, 1H).

Step G: Preparation of Ethyl3-(2-Ethoxy-2-oxoethyl)-7-methoxy-1,2,3,4-tetrahydroeyelopenta[β]indole-3-carboxylate

To a suspension of (4-methoxyphenyl)hydrazine hydrochloride (379.5 g,2.17 mol) and ethyl 1-(2-ethoxy-2-oxoethyl)-2-oxocyclopentanecarboxylate(526 g, 2.17 mol) in EtOH (2.0 L), AcOH (131 g, 124 mL, 2.17 mol) wasadded and the mixture was stirred at 75° C. for 18 h under N₂. The finedark brown suspension was allowed to cool and neutralized with saturatedaqueous NaHCO₃. The solvent was evaporated under reduced pressure. Thebrown oily residue was taken up in EtOAc (2 L), filtered and theorganics were washed with water (3×500 mL) and brine (2×500 mL). Thecombined aqueous layers were re-extracted with EtOAc. The combinedorganics were dried (MgSO₄) and the solvent was evaporated under reducedpressure to give the title compound (703.4 g) as a thick dark brown oil.LCMS m/z=346.2 [M+H]⁺; ¹H NMR (400 MHz, DMSO-d₆) δ ppm 1.15 (t, J=7.2Hz, 3H), 1.17 (t, J=7.2 Hz, 3H), 2.48-2.42 (m, 1H), 2.81 (d, J=16.6 Hz,1H), 2.82-2.70 (m, 2H), 3.05-2.99 (m, 1H), 3.18 (d, J=16.6 Hz, 1H), 3.73(s, 3H), 4.12-4.00 (m, 4H), 6.67 (dd, J=8.8, 2.5 Hz, 1H), 6.85 (d, J=2.4Hz, 1H), 7.21 (d, J=8.7 Hz, 1H), 10.57 (s, 1H).

Step H: Preparation of3-(Carboxymethyl)-7-methoxy-1,2,3,4-tetrahydrocyclopenta[β]indole-3-carboxylicAcid

A 50 wt % aqueous solution of NaOH (346 g, 4.32 mol, 4 equiv.) wasslowly added to a solution of ethyl3-(2-ethoxy-2-oxoethyl)-7-methoxy-1,2,3,4-tetrahydrocyclopenta[β]indole-3-carboxylate(373 g, 1.08 mol) in EtOH (2.0 L) and the resulting mixture was stirredat 60° C. for 18 h under N₂. The brown suspension was neutralized at 0°C. with 6 N HCl and the solvent was evaporated. The brown residue waspartitioned between H₂O (2 L) and EtOAc (1 L) and the layers separated.The aqueous layer was further washed with EtOAc (3×500 mL) and the pH ofthe aqueous phase was adjusted to 3-4 with 6 N HCl. The precipitate wascollected and dried under vacuum at ambient temperature overnight togive the title compound (191.4 g) as a brown solid. LCMS m/z=290.4[M+H]⁺; ¹H NMR (400 MHz, DMSO-d₆) δ ppm 2.43-2.36 (m, 1H), 2.68 (d,J=16.9 Hz, 1H), 2.82-2.69 (m, 2H), 3.07-3.01 (m, 1H), 3.12 (d, J=16.9Hz, 1H), 3.72 (s, 3H), 6.66 (dd, J=8.7, 2.4 Hz, 1H), 6.84 (d, J=2.4 Hz,1H), 7.20 (d, J=8.6 Hz, 1H), 10.55 (s, 1H), 12.30 (s, 2H).

Step I: Preparation of2-(7-Methoxy-1,2,3,4-tetrahydrocyclopenta[β]indol-3-yl)acetic Acid

A solution of3-(carboxymethyl)-7-methoxy-1,2,3,4-tetrahydrocyclopenta[β]indole-3-carboxylicacid (191 g, 0.66 mol) in AcOH (1.0 L) was stirred at 60° C. for 4.5 hunder N₂. The dark brown solution was concentrated. The precipitate wascollected, washed with H₂O (3×500 mL) and dried at 40° C. under vacuumovernight to give the title compound (126.4 g) as a brown solid. LCMSm/z=246.1 [M+H]⁺; ¹H NMR (400 MHz, DMSO-d₆) δ ppm 2.12-2.04 (m, 1H),2.35 (dd, 16.0, 9.1 Hz, 1H), 2.77-2.60 (m, 4H), 3.50-3.43 (m, 1H), 3.72(s, 3H), 6.62 (dd, J=8.8, 2.5 Hz, 1H), 6.81 (d, J=2.3 Hz, 1H), 7.18 (d,J=8.7 Hz, 1H), 10.42 (s, 1H), 12.16 (s, 1H).

Step J: Preparation of Ethyl2-(7-Hydroxy-1,2,3,4-tetrahydrocyclopenta[β]indol-3-yl)acetate

To a solution of BBr₃ (115 g, 43.3 mL, 458 mmol, 3 equiv.) in CH₂Cl₂ (70mL) a suspension of2-(7-methoxy-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetic acid(37.44 g, 153 mmol) in CH₂Cl₂ (300 mL) was added slowly whilemaintaining the reaction temperature between −5° C. to 0° C. Theresulting dark brown suspension was stirred at −5 to 0° C. for anadditional 1 h. EtOH (187 mL) was added dropwise to the reaction mixturewhile maintaining the temperature between 0-10° C. The resultingsolution was heated at 40° C. for 30 min. The solution was cooled andthe pH adjusted to 8 by adding 10 N NaOH (142.9 mL, 1.43 mol) slowlywhile maintaining the temperature between 0-3° C. The solvent wasremoved under reduced pressure until about 200 mL of concentrateremained. The pH was adjusted to about 7 with concentrated HCl, thesuspension filtered, the solids washed with H₂O (3×200 mL) and driedunder vacuum at ambient temperature overnight. The light brown materialwas dissolved in EtOAc (200 mL), and filtered washing the solids withEtOAc. The combined organics were washed with saturated aqueous NaHCO₃(2×200 mL), brine (200 mL), dried (Na₂SO₄) and the solvent rotaryevaporated to give the title compound (35.2 g) as a light brown solid.LCMS m/z=260.1 [M+H]⁺; ¹H NMR (400 MHz, DMSO-d₆) δ ppm 1.20 (t, J=7.1Hz, 3H), 2.11-2.03 (m, 1H), 2.42 (dd, J=15.7, 8.9 Hz, 1H), 2.71-2.55 (m,3H), 2.76 (dd, J=15.7, 5.5 Hz, 1H), 3.49-3.42 (m, 1H), 4.11 (q, J=7.1Hz, 2H), 6.49 (dd, J=8.6, 2.3 Hz, 1H), 6.62 (d, J=2.1 Hz, 1H), 7.07 (d,J=8.6 Hz, 1H), 8.47 (s, 1H), 10.25 (s, 1H).

Step K: Preparation of Ethyl2-(7-(4-Cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetate

To a solution of4-(chloromethyl)-1-cyclopentyl-2-(trifluoromethyl)benzene (46.2 g, 176mmol, 1.2 eq) in DMF (400 mL) ethyl2-(7-hydroxy-1,2,3,4-tetrahydrocyclopenta[O]indol-3-yl)acetate (38.0 g,147 mmol) was added in one portion followed by Cs₂CO₃ (71.6 g, 220 mmol,1.5 eq). An exotherm was observed over the first 15 min (temperatureincreased to 72.8° C.) after which the mixture was further stirred at50° C. under N₂ for 13.5 h. The reaction mixture was allowed to cool,filtered under suction washing the solids with EtOAc and the filtrateevaporated under reduced pressure. The dark brown oily residue was takenup in EtOAc, washed with H₂O (3×300 mL) and the aqueous phasere-extracted with EtOAc. The combined organics were dried (MgSO₄),filtered and rotary evaporated to give the title compound (77 g) as athick dark brown oil which was used in the next step without furtherpurification. LCMS m/z=486.4 [M+H]⁺; ¹H NMR (400 MHz, DMSO-d₆) δ ppm1.19 (t, J=7.2 Hz, 3H), 1.73-1.54 (m, 4H), 1.89-1.77 (m, 2H), 2.12-1.94(m, 3H), 2.44 (dd, J=17.2, 8.4 Hz, 1H), 2.81-2.60 (m, 4H), 3.30-3.20 (m,1H), 3.54-3.44 (m, 1H), 4.12 (q, J=7.2 Hz, 2H), 5.12 (s, 2H), 6.72 (dd,J=8.8, 2.4 Hz, 1H), 6.94 (d, J=2.4 Hz, 1H), 7.20 (d, J=8.8 Hz, 1H), 7.62(d, J=7.8 Hz, 1H), 7.73-7.67 (m, 2H), 10.47 (s, 1H).

Step L: Preparation of2-(7-(4-Cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)aceticAcid

To a solution of ethyl2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)acetate(71.2 g, 147 mmol) in 1,4-dioxane (400 mL) an aqueous solution ofLiOH.H₂O (220 mL, 2 M, 3 eq) was added. The resulting two phase mixturewas stirred at 50° C. under N₂ for 5 h. The reaction mixture was allowedto cool and the pH adjusted to 3-4 with 6 N HCl. The solvent was rotaryevaporated and to the two phase aqueous/product mixture CH₂Cl₂ added.The layers were separated and the organics washed with H₂O (2×300 mL).The combined aqueous phases were re-extracted with CH₂Cl₂. The combinedorganic layers were dried (MgSO₄), filtered and rotary evaporated. Thedark brown oily residue was taken up in MeOH and the solvent evaporatedunder reduced pressure. The dark brown residue was taken up again in aminimum amount of MeOH and left in the fridge over 16 h. The precipitatewas collected under suction, washing the solids with hexanes, and driedunder high vacuum to afford the product (34.5 g, 51%) as an off-whitesolid. The filtrate containing product was concentrated to dryness togive a dark brown fluffy solid (33.6 g) which was processed furtherseparately. LCMS m/z=458.2 [M+H]⁺; ¹H NMR (400 MHz, DMSO-d₆) δ ppm1.76-1.57 (m, 4H), 1.93-1.81 (m, 2H), 2.16-1.97 (m, 1H), 2.38 (dd,J=15.2, 8.8 Hz, 1H), 2.80-2.62 (m, 4H), 3.32-3.23 (m, 1H), 3.54-3.44 (m,1H), 5.15 (s, 2H), 6.74 (dd, J=8.8, 2.4 Hz, 1H), 6.96 (d, J=2.4 Hz, 1H),7.23 (d, J=8.8 Hz, 1H), 7.65 (d, J=8.4 Hz, 1H), 7.76-7.70 (m, 2H), 10.48(s, 1H), 12.20 (s, 1H).

Example 1.31 Preparation of2-(7-(4-Cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)aceticAcid Step A: Preparation of 1-Cyclopentyl-2-(trifluoromethyl)benzene

In a 5 L 3-necked, round-bottomed flask fitted with a mechanicalstirrer, a temperature probe, a dry nitrogen inlet, a condenser and anaddition funnel was placed anhydrous THF (750 mL) and magnesium (40.5 g,1667 mmol) under N₂. The slurry was cooled to 10° C. in an ice bath. Asolution of FeCl₃ (18.02 g, 111 mmol) in anhydrous THF (80 mL) (Note:dissolution of FeCl₃ in THF was exothermic) was added dropwise to themagnesium slurry via a syringe.N1,N1,N2,N2-tetramethylethane-1,2-diamine (201 mL, 1333 mmol) was addedto the reaction mixture via a 500 mL addition funnel at 15° C. causingthe temperature to go up to 22.5° C. The reaction mixture was cooled to18° C. and stirred at that temperature for 1 h and 45 min. It was thenheated to 44-45° C., stirred for 1 h, cooled to 5-10° C., and added amixture of 1-bromo-2-(trifluoromethyl)benzene (150 mL, 1111 mmol) andbromocyclopentane (143 mL, 1333 mmol) dropwise while keeping internaltemperature under 30° C. (temperature was maintained between 22 to 30°C.). After addition was complete, the reaction mixture was cooled to17-18° C. and stirred overnight at room temperature. This main reactionmixture was cooled to 10° C. and added magnesium (15 g). Meanwhile, in aseparate 1 L round-bottomed flask under N₂, Mg (20 g, 0.5 eq) in THF(300 mL) was added a solution of FeCl₃ (9 g, 0.5 eq) in anhydrous THF(30 mL) like described above. The obtained mixture was stirred at roomtemperature for 30 min, heated to 45° C. for 1 h, cooled to roomtemperature, and added dropwise into the main reaction mixture via anaddition funnel (remaining magnesium was transferred by a spatula) whilekeeping the internal temperature under 30° C. The reaction was continuedat room temperature for 1 h, cooled to 5° C. (ice bath) and quenchedslowly with saturated NH₄Cl solution (150 mL) (quench was exothermicsatd. NH4Cl was added slowly with efficient stirring). Celite was addedafter quenching the reaction and stirred well. The mixture was filteredthrough a 3 L sintered funnel. The filter cake was washed with THF. Thefiltrate was concentrated under reduced pressure at 37° C. (bathtemperature)/155 Torr to obtain a brown oil. The oil was cooled by icebath and 6N HCl (500 mL) was poured into it slowly with efficientstirring) (addition of HCl was exothermic initially, then the exothermsubsided). The mixture was extracted with hexane (2×400 mL). The hexanelayer was separated out and filtered through a pad of celite. Thefiltrate (hexane layer) was washed with water (3×300 mL), dried (Na₂SO₄)and silica (550 g) added; slurried well. The slurry was filtered and thefiltrate (light yellow in color) was concentrated under reduced pressure(rotavapor; bath temp. 37 C at 185-188 Torr), to give the title compoundas a light orange oil, (190 g, 91.4% purity by LC at 214 nm). ¹H NMR(400 MHz, DMSO-d₆) δ 1.56-1.71 (m, 4H), 1.80-1.88 (m, 2H), 1.96-2.05 (m,2H), 3.22-3.29 (m, 1H), 7.35-7.40 (m, 1H), 7.16-7.65 (m, 2H).

Step B: Preparation of4-(Chloromethyl)-1-cyclopentyl-2-(trifluoromethyl)benzene

In a 1 L, 3-necked reaction flask fitted with a mechanical stirrer, atemperature probe, an addition funnel and a dry nitrogen inlet wasplaced 1-cyclopentyl-2-(trifluoromethyl)benzene (50 g, 233 mmol). Thematerial was stirred and cooled to −12° C. (dry ice/IPA bath).Concentrated sulfuric acid (100 mL, 1877 mmol) was added dropwise sothat the temperature was maintained between −12° C. to −10° C. Themixture was cooled to −15° C. and s-trioxane (27.3 g, 303 mmol) wasadded in 3 batches (9.1 g each batch) while the temperature wasmaintained at between −15° C. to −10° C. The mixture was stirred at −10°C. and almost immediately sulfurochloridic acid (28.1 mL, 420 mmol) wasadded slowly maintaining the temperature at between −10° C. to −5° C.The mixture was stirred for 20 min at −5° C. and 3 h between −2 to −3°C. The reaction mixture was slowly poured (with efficient stirring) intoice-water (1 L). MTBE (700 mL) was added and the mixture was stirredwell. Celite (300 g) was added and stirred well. The celite slurry wasfiltered and the celite bed was washed with MTBE. The aqueous layer ofthe filtrate was separated and extracted with MTBE (1×700 mL). Thecombined MTBE layer was washed with water (1×500 mL) followed bysaturated NaHCO₃ (2×350 mL). The MTBE layer was then washed with water(2×500 mL), dried (Na₂SO₄) and filtered. The filtrate was concentrated(at 38° C., bath temperature; 200 Torr) to give a yellow oil. The oilwas taken up in hexane (500 mL) and filtered through a bed of silica;then the silica bed was washed with hexane. The filtrate wasconcentrated under reduced pressure (38° C., bath temperature; at 200Torr) to give the title compound as a light yellow oil (36.2 g; 89%purity by LC at 214 nm). ¹H NMR (400 MHz, DMSO-d₆) δ 1.55-1.72 (m, 4H),1.78-1.89 (m, 2H), 1.94-2.04 (m, 2H), 3.19-3.28 (m, 1H), 4.82 (s, 2H),7.62-7.72 (m, 3H).

Step C: Preparation of1-Cyclopentyl-4-((4-nitrophenoxy)methyl)-2-(trifluoromethyl)benzene

In a 1 L flask equipped with a stirrer, thermocouple, condenser and anitrogen inlet, was placed nitrophenol (28 g, 201 mmol) in DMA (150 mL)and potassium carbonate powder (28.7 g, 207 mmol).4-(Chloromethyl)-1-cyclopentyl-2-(trifluoromethyl)benzene (45.4 g, 173mmol) was added and washed in with DMA (120 mL). The reaction was heatedat 80° C. (bath, internal 77° C.) overnight. The mixture was cooled andpoured into ice water (1 L). The solids formed were allowed to settlewith stirring for 4 h and collected by filtration. The solid collectedwas stirred in sodium bicarbonate solution (300 mL), filtered, washedwith water, and air dried. The pale yellow residue was washed withhexanes (250 mL) and the solids were dried in vacuum oven overnight togive the title compound (41.4 g, ˜85% pure by LC). LCMS m/z=366.2[M+H]⁺.

Step D: Preparation of4-(4-Cyclopentyl-3-(trifluoromethyl)benzyloxy)aniline hydrochloride

In a 2 L flask equipped with stirrer and thermocouple, was placed1-cyclopentyl-4-((4-nitrophenoxy)methyl)-2-(trifluoromethyl)benzene (40g, 109 mmol) in ACN (520 mL). Ammonium chloride (3M, 520 mL) was addedand the mixture was stirred and cooled to 2.5° C. Zinc (35.8 g, 547mmol) was added in portions keeping temperature below 5° C. Afteraddition was completed, the reaction mixture was allowed to warm to roomtemperature and stirred overnight. The mixture was filtered through abed of celite (50 g) and the filter bed was washed with ACN (150 mL).The aqueous layer of the filtrate was separated and back extracted withisopropyl acetate (200 mL). The combined organic layers were dried oversodium sulfate (50 g), filtered and concentrated. The residue wasdissolved in ethanol (120 mL), added HCl (1.25 M in EtOH, 140 mL), andstirred at ambient for 2.5 hours. After removal of the solvent, theresidual solids was triturated with ACN (120 mL), filtered, washed withACN (2×50 mL), and dried under vacuum to give the title compound (29.8g).

Step E: Preparation of(4-(4-Cyclopentyl-3-(trifluoromethyl)benzyloxy)phenyl)hydrazineHydrochloride

4-(4-Cyclopentyl-3-(trifluoromethyl)benzyloxy)aniline hydrochloride (30g, 81 mmol) was suspended in water (285 mL), and concentrated HCl (18mL) was added. The suspension was stirred efficiently and cooled inice/IPA bath to −0° C. Sodium nitrite (5.57 g, 81 mmol) in water (12 mL)was added. After addition, the reaction was stirred at 2° C. for 40minutes. Some solids on the side were washed with ACN (10 mL). Themixture was cooled to −1° C. and tin (II)chloride (45.9 g, 242 mmol)dissolved in concentrated HCl (30 mL) was added slowly. A thickprecipitate was formed and stirring was continued for 30 minutes. Themixture was warmed to room temperature and stirred for 3 h. The mixturewas filtered, washed with HCl (0.1 M) and the solid was dried undervacuum to give the title compound (40.6 g).

Step F: Preparation of Ethyl7-(4-Cyclopentyl-3-(trifluoromethyl)benzyloxy)-3-(2-ethoxy-2-oxoethyl)-1,2,3,4-tetrahydrocyclopenta[b]indole-3-carboxylate

In a 1 L flask was placed EtOH (500 mL). Sulfuric acid (2.4 g, 23.98mmol) was added at 40° C., followed by ethyl1-(2-ethoxy-2-oxoethyl)-2-oxocyclopentanecarboxylate (15.2 g, 62.7mmol). (4-(4-Cyclopentyl-3-(trifluoromethyl)benzyloxy)phenyl)hydrazinehydrochloride (24.0 g, 62.0 mmol) was added and the solution becamelight yellow and homogenous. The reaction mixture was refluxed overnightwith a Dean-Starks condenser attached. The mixture was cooled andextracted in ethyl acetate (3×100 mL). The organics were washed withwater (200 mL), sodium bicarbonate solution (2×70 mL), water (100 mL),dried over magnesium sulfate, and concentrated. The residue wasdissolved in hexanes/ethyl acetate (80:20, 300 mL), added silica gel (30g) and stirred for 35 minutes. The slurry was filtered, washed with thesame elution solvent (100 mL) and the filtrate was concentrated to givethe title compound (26.7 g). LCMS m/z=558.5 [M+H]⁺.

Step G: Preparation of Sodium3-(Carboxylatomethyl)-7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indole-3-carboxylate

In a 500 mL, flask was placed ethyl7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-3-(2-ethoxy-2-oxoethyl)-1,2,3,4-tetrahydrocyclopenta[b]indole-3-carboxylate(24.1 g, 43.2 mmol) in isopropanol (275 mL). Sodium hydroxide solution(20%, 129.5 mL, 130 mmol) was added and the mixture was heated at 100°C. (bath) for 2.5 h. The mixture was cooled, filtered, washed withisopropanol, and dried overnight under vacuum at 40° C. to give thetitle compound (17.5 g). LCMS m/z=502.6 [M−2Na+3H]⁺.

Step H: Preparation of2-(7-(4-Cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)aceticAcid

To a stirred solution of sodium3-(carboxylatomethyl)-7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indole-3-carboxylate(16.5 g, 30.2 mmol) in water at 40° C. was added ammonium chloridesolution (9.71%, 100 mL). The reaction was heated at 92° C. (bath) for4.4 h. The mixture was refrigerated overnight, decanted and trituratedwith ice cold 6N HCl (100 mL). The solid was collected by filtration,washed with diluted HCl (100 mL) and dried overnight in a vacuum oven at40° C. to give the title compound (8.5 g). LCMS m/z=458.3 [M+H]⁺.

Example 1.32 Preparation of the Ca salt of the 2^(nd) Enantiomer ofCompound 12

Prior to use, the 2^(nd) enantiomer of Compound 12, as described inExample 1.29, was slurried in acetonitrile overnight, filtered and driedto produce a crystalline form. To the crystalline form (40 mg) was addedacetonitrile (1 mL) and the mixture was warmed to 60° C. The counterionwas added by adding 20 μL of calcium acetate solution (2 M) and 20 μL,of water then seeding with crystalline salt and allowing to slowly coolto room temperature. The resulting solid was filtered and dried to givea white solid.

Example 1.33 Preparation of L-Arginine Salt of the 2^(nd) Enantiomer ofCompound 12

The 2^(nd) enantiomer of Compound 12 as described in Example 1.29 (174.7mg, 0.381 mmol) was dissolved in IPA (1.57 mL) and L-arginine (66.4 mg,0.381 mmol) was added as a solution in water (263 μL). The homogeneoussolution was warmed to 40° C. After 15 min. at this temperature, aprecipitate had formed. The reaction mixture was warmed to 70° C.causing the precipitate to dissolve. The heat bath was turned off. Aprecipitate began to form at 40° C. and the reaction was allowed to coolto 28° C. before collecting the solids by filtration. The solids werewashed with 14% water in IPA to give the L-arginine salt of the titlecompound (130 mg).

Example 1.34 Preparation of the D-Lysine salt of the 1^(st) Enantiomerof Compound 12

To the 1^(st) enantiomer of Compound 12 as described in Example 1.29 inacetonitrile with 3% water was added D-lysine (1 M aqueous solution).After stirring overnight at ambient temperature, the resulting solid wasfiltered and dried.

Example 1.35 Preparation of the (R)-1-phenethylamine salt acetonitrilesolvate of the 2^(nd) Enantiomer of Compound 12

To a solution of the 2^(nd) enantiomer of Compound 12 as described inExample 1.29 in acetonitrile was added (R)-1-phenethylamine (1equivalent). The mixture was heated briefly and allowed to cool down. Aprecipitate was formed, filtered and dried. Single crystals of the(R)-1-phenylethanamine salt of the 2^(nd) enantiomer of Compound 12 (asdescribed in Example 1.29) were recrystallized by slow evaporation fromacetonitrile and acetone and subjected to X-ray crystallographyanalysis. An acetonitrile solvate was observed in the ratio of 4 saltmoieties to 1 acetonitrile molecule.

Example 2 Homogeneous Time-Resolved Fluorescence (HTRF®) Assay ForDirect cAMP Measurement

Compounds were screened for agonists of the S1P1 receptor (e.g., humanS1P1 receptor) using the HTRF® assay for direct cAMP measurement(Gabriel et al., Assay and Drug Development Technologies, 1:291-303,2003) and recombinant CHO-K1 cells stably transfected with S1P1receptors. CHO-K1 cells were obtained from ATCC® (Manassas, Va.; Catalog# CCL-61). An agonist of the S1P1 receptor was detected in the HTRF®assay for direct cAMP measurement as a compound which decreased cAMPconcentration. HTRF® assay also was used to determine EC₅₀ values forS1P1 receptor agonists.

Principle of the Assay:

HTRF® assay kit was purchased from Cisbio-US, Inc. (Bedford, Mass.;Catalog #62AM4PEC). The HTRF® assay supported by the kit is acompetitive immunoassay between endogenous cAMP produced by the CHO-K1cells and tracer cAMP labeled with the dye d2. The tracer binding isvisualized by a monoclonal anti-cAMP antibody labeled with Cryptate. Thespecific signal (i.e., fluorescence resonance energy transfer, FRET) isinversely proportional to the concentration of unlabeled cAMP in thestandard or sample.

Standard Curve:

The fluorescence ratio (665 nm/620 nm) of the standards (0.17 to 712 nMcAMP) included in the assay was calculated and used to generate a cAMPstandard curve according to the kit manufacturer's instructions. Thefluorescence ratio of the samples (test compound or compound buffer) wascalculated and used to deduce respective cAMP concentrations byreference to the cAMP standard curve.

Setup of the Assay:

The HTRF® assay was carried out using a two-step protocol essentiallyaccording to the kit manufacturer's instructions, in 20 μL total volumeper well in 384-well plate format (ProxiPlates; PerkinElmer, Fremont,Calif.; catalog #6008280). To each of the experimental wells wastransferred 1500 recombinant CHO-K1 cells in 5 μL phosphate bufferedsaline containing calcium chloride and magnesium chloride (“PBS+”;Invitrogen, Carlsbad, Calif.; catalog #14040) supplemented with IBMX(250 μM) and rolipram (20 μM) (phosphodiesterase inhibitors;Sigma-Aldrich, St. Louis, Mo.; catalog #15879 and catalog # R6520,respectively), followed by test compound in 5 μL compound buffer(PBS+supplemented with 10 μL NKH477 (water-soluble forskolin derivative;SignaGen Laboratories, Gaithersburg, Md.; catalog # PKI-NKH477-010)) or5 μL compound buffer. The plate was then incubated at room temperaturefor 1 h. To each well was then added 5 μL cAMP-d2 conjugate in lysisbuffer and 5 μL Cryptate conjugate in lysis buffer according to the kitmanufacturer's instructions. The plate was then further incubated atroom temperature for 1 hour, after which the assay plate was read.

Assay Readout:

HTRF® readout was accomplished using a PHERAstar (BMG LABTECH Inc.,Durham, N.C.) or EnVision™ (PerkinElmer, Fremont Calif.) microplatereader.

Certain compounds of the present invention and their correspondingactivity values are shown in Table B.

TABLE B Compound No. EC₅₀ S1P1 (HTRF) 4 16 nM 8  9 nM 10 26 nM

Certain other compounds of the invention had activity values rangingfrom about 35 pm to about 362 nM in this assay.

Example 3 Cellular/Functional Ca²⁺ Assay for Agonist Activity on S1P3Receptor

A compound of the invention can be shown to have no or substantially noagonist activity on the S1P3 receptor by using in assay a humanneuroblastoma cell line which endogenously expresses S1P3(predominantly), S1P2 and S1P5 receptors, but not S1P1 or S1P4receptors, based on mRNA analysis (Villullas et al., J. Neurosci. Res.,73:215-226, 2003). Of these, S1P3 and S1P2 receptors respond toagonists, such as S1P, with an intracellular calcium increase. No orsubstantially no increase of intracellular calcium in response to a testcompound is indicative of the test compound exhibiting no orsubstantially no agonist activity on the S1P3 receptor. Such an assaycan be performed commercially, e.g. by Caliper LifeSciences (Hopkinton,Mass.).

Assay:

The human neuroblastoma cells are washed and resuspended inphysiological buffer. The cells are then loaded with dye that measuresintracellular calcium. S1P is used as a reference agonist. Afteraddition of S1P or a test compound, fluorescence is measured at 485 nmexcitation/525 nm emission every 2 s for at least 60 s. Calciumionophore A23187 is then added as an internal positive control

Example 4 Effect of Compounds in Peripheral Lymphocyte Lowering (PLL)Assay

A compound of the invention can be shown to induce peripheral lymphocytelowering (PLL).

A. Mouse PLL Assay.

Animals:

Male BALB/c mice (Charles River Laboratories, Wilmington, Mass.) werehoused four per cage and maintained in a humidity-controlled (40 to 60%)and temperature-controlled (68 to 72° F.) facility on a 12 h:12 hlight/dark cycle (lights on at 6:30 am) with free access to food (HarlanTeklad, Orange, Calif., Rodent Diet 8604) and water. Mice were allowedone week of habituation to the animal facility before testing.

PLL Assay:

Mice were given an oral dose of Compound 5, Compound 7 or dosing vehicle(0.5% methylcellulose) in a total volume of 10 mL/kg. Peripheral bloodsamples were collected at 5 hours post-dose. The mice were anesthetizedwith isoflurane and blood was collected via cardiac puncture. A completecell count (CBC), including lymphocyte count, was obtained using aCELL-DYN® 3700 (Abbott Laboratories, Abbott Park, Ill.) instrument.Results are presented in FIGS. 1 and 2, in which peripheral bloodlymphocyte (PBL) count is shown for the 5 hour group. Reduction of thePBL count by the test compound in comparison with vehicle is indicativeof the test compound exhibiting activity or inducing peripherallymphocyte lowering. It is apparent from inspection of FIGS. 1 and 2that Compound 5 and Compound 7 exhibited activity for inducing PBLlowering (lymphopenia) in the mouse.

PLL Assay:

Mice were given a 1.00 mg/kg oral dose of the 2^(nd) enantiomer ofcompound 12 (isolated after resolution of compound 12 by HPLC, with aretention time of 13.9 min per the conditions reported in Example 1.29)or dosing vehicle (0.5% methylcellulose in sterile water) in a totalvolume of 10 mL/kg. Peripheral blood samples were collected at 5 hourspost-dose. The mice were anesthetized with isoflurane and blood wascollected via cardiac puncture. A complete cell count (CBC), includinglymphocyte count, was obtained using a CELL-DYN® 3700 (AbbottLaboratories, Abbott Park, Ill.) instrument. Results are presented inFIG. 7, in which peripheral blood lymphocyte (PBL) count is shown forthe 5 hour group. Reduction of the PBL count by the test compound incomparison with vehicle is indicative of the test compound exhibitingactivity or inducing peripheral lymphocyte lowering. It is apparent frominspection of FIG. 7 that the 2^(nd) enantiomer of compound 12 (isolatedafter resolution of compound 12 by HPLC, with a retention time of 13.9min per the conditions reported in Example 1.29) exhibited activity forinducing PBL lowering (lymphopenia) in the mouse.

B. Rat PLL Assay.

Animals:

Male Sprague-Dawley rats (7 weeks of age at start of study) (CharlesRiver Laboratories) were housed two per cage and maintained in ahumidity-controlled (40-60%) and temperature-controlled (68-72° F.)facility on a 12 h:12 h light/dark cycle (lights on at 6:30 am) withfree access to food (Harlan Teklad, Orange, Calif., Rodent Diet 8604)and water. Rats were allowed one week of habituation to the animalfacility before testing.

PLL Assay:

Rats were given a 1.00 mg/kg oral dose of the 2^(nd) enantiomer ofcompound 12 (isolated after resolution of compound 12 by HPLC, with aretention time of 13.9 min per the conditions reported in Example 1.29)or dosing vehicle (0.5% methylcellulose in sterile water) in a totalvolume of 1.00 mL/kg. Peripheral blood samples were collected at 5 hourspost-dose. Blood was collected via indwelling catheter. A complete cellcount (CBC), including lymphocyte count, was obtained using a CELL-DYN®3700 (Abbott Laboratories, Abbott Park, Ill.) instrument. Results arepresented in FIG. 8, in which peripheral blood lymphocyte (PBL) count isshown for the 5 hour group. Reduction of the PBL count by the testcompound in comparison with vehicle is indicative of the test compoundexhibiting activity or inducing peripheral lymphocyte lowering. It isapparent from inspection of FIG. 8 that the 2^(nd) enantiomer ofcompound 12 (isolated after resolution of compound 12 by HPLC, with aretention time of 13.9 min per the conditions reported in Example 1.29)exhibited activity for inducing PBL lowering (lymphopenia) in the rat.

Example 5 Effect of Compounds on Experimental AutoimmuneEncephalomyelitis (EAE)

A compound of the invention can be shown to have therapeutic efficacy inmultiple sclerosis by showing it to have therapeutic efficacy inexperimental autoimmune encephalomyelitis (EAE), an animal model formultiple sclerosis. In certain exemplary well-established models, EAE isinduced in rodents by injection of myelin oligodendrocyte glycoprotein(MOG) peptide, by injection of myelin basic protein (MBP) or byinjection of proteolipid protein (PLP) peptide.

A. MOG-Induced EAE in Mice.

Animals:

Female C57BL/6 mice (8 to 10 weeks of age at start of study) (JacksonLaboratory, Bar Harbor, Me.) were housed four per cage and maintained ina humidity-controlled (40-60%) and temperature-controlled (68-72° F.)facility on a 12 h:12 h light/dark cycle (lights on at 6:30 am) withfree access to food (Harlan Teklad, Orange, Calif., Rodent Diet 8604)and water. Mice were allowed one week of habituation to the animalfacility before testing.

Induction of EAE:

Mice were immunized subcutaneously, 50 μL per hind flank, with a totalof 100 μg MOG₃₅₋₅₅ peptide emulsified 1:1 with Complete Freund'sadjuvant containing 4 mg/mL heat-killed Mycobacterium tuberculosis. Micealso received 200 ng pertussis toxin intraperitoneally on the day ofimmunization and 48 h later.

Clinical Scoring:

Severity of disease symptoms was scored as follows (in increasing orderof severity): 0=normal; 1=limp tail OR hind limb weakness; 2=limp tailAND limb weakness/weakness of 2 or more limbs; 3=severe limb weakness orsingle limb paralysis; 4=paralysis of 2 or more limbs; 5=death.

Drug Treatment:

Mice were dosed orally, with vehicle or the 2^(nd) enantiomer ofcompound 12 (isolated after resolution of compound 12 by HPLC, with aretention time of 13.9 min per the conditions reported in Example 1.29),once a day from day 3 until day 21. Dosing volume is 5 mL/kg. The 2^(nd)enantiomer of compound 12 (isolated after resolution of compound 12 byHPLC, with a retention time of 13.9 min per the conditions reported inExample 129) was dosed at 0.3 mg/kg, 1 mg/kg and 3 mg/kg. Mice wereweighed daily. Mice were monitored daily from day 7 onward for diseasesymptoms. After the last dose on day 21, disease progression wasmonitored daily for 2 more weeks. Reduction of the severity of diseasesymptoms by the 2^(nd) enantiomer of compound 12 (isolated afterresolution of compound 12 by HPLC, with a retention time of 13.9 min perthe conditions reported in Example 1.29) in comparison with vehicle wasindicative of the test compound exhibiting therapeutic efficacy in EAE.It is apparent from inspection of FIG. 10 that the 2^(nd) enantiomer ofcompound 12 (isolated after resolution of compound 12 by HPLC, with aretention time of 13.9 min per the conditions reported in Example 1.29)exhibited activity in the mouse EAE assay.

B. PLP-Induced EAE in Mice.

Animals:

Female SJL/J mice (8 to 10 weeks of age at start of study) (JacksonLaboratory, Bar Harbor, Me.) are housed four per cage and maintained ina humidity-controlled (40-60%) and temperature-controlled (68-72° F.)facility on a 12 h:12 h light/dark cycle (lights on at 6:30 am) withfree access to food (Harlan-Teklad Western Res, Orange, Calif., RodentDiet 8604) and water. Mice are allowed one week of habituation to theanimal facility before testing.

Induction of EAE:

Mice are immunized subcutaneously with 100 μg PLP₁₃₉₋₁₅₁ peptideemulsified 1:1 with Complete Freund's adjuvant containing 4 mg/mLheat-killed Mycobacterium tuberculosis. Mice also receive 200 ngpertussis toxin intravenously on the day of immunization.

Clinical Scoring:

Severity of disease symptoms is scored as follows (in increasing orderof severity): 0=normal; 1=limp tail OR hind limb weakness; 2=limp tailAND limb weakness/weakness of 2 or more limbs; 3=severe limb weakness orsingle limb paralysis; 4=paralysis of 2 or more limbs; 5=death.

Drug Treatment:

Mice are dosed orally, with vehicle or a test compound, once a day fromday 3 until day 21. Dosing volume is 5 ml/kg. The test compound is dosedat, e.g., 1 mg/kg, 3 mg/kg, 10 mg/kg or 30 mg/kg. Mice are weigheddaily. Mice are monitored daily from day 7 onward for disease symptoms.After the last dose on day 21, disease progression is monitored dailyfor two more weeks.

C. MBP-Induced EAE in Rats.

Animals:

Male Lewis rats (325-375 g at start of study) (Harlan, San Diego,Calif.) are housed two per cage and maintained in a humidity-controlled(30-70%) and temperature-controlled (20-22° C.) facility on a 12 h:12 hlight/dark cycle (lights on at 6:30 A.M.) with free access to food(Harlan-Teklad Western Res., Orange, Calif., Rodent Diet 8604) andwater. Rats are allowed one week of habituation to the animal facilitybefore testing. During the study, rats are weighed daily prior toclinical scoring at 11 am.

Induction of EAE:

Myelin basic protein (MBP; guinea pig) is dissolved in sterile saline ata concentration of 1 mg/ml, and then emulsified 1:1 with CompleteFreund's adjuvant (1 mg/ml). 50 μL of this emulsion is administered byintraplantar (ipl) injection into both hind paws of each rat, for atotal injected volume of 100 μL per rat and a total dose of 50 μg of MBPper rat.

Clinical Scoring:

Severity of disease symptoms is scored daily after body weighing andbefore drug dosing. Severity of disease symptoms is scored as follows(in increasing order of severity): 0=normal; 1=tail OR limb weakness;2=tail AND limb weakness; 3=severe hind limb weakness or single limbparalysis; 4=loss of tail tone and paralysis of 2 or more limbs;5=death.

Drug Treatment:

Rats are dosed orally, with vehicle or a test compound, 1 hour prior toMBP injection on day 0 and daily thereafter, after clinical scoring, forthe duration of the study. Dosing volume is 5 mL/kg. The test compoundis dosed at, e.g., 1 mg/kg, 3 mg/kg, 10 mg/kg or 30 mg/kg. Reduction ofthe severity of disease symptoms by the test compound in comparison withvehicle is indicative of the test compound exhibiting therapeuticefficacy in EAE.

Example 6 Effect of Compounds on Type I Diabetes

A compound of the invention can be shown to have therapeutic efficacy intype I diabetes using an animal model for type I diabetes, such ascyclophosphamide-induced type I diabetes in mice.

Animals:

Baseline blood glucose measurements are taken from 9-10 week old femaleNOD/Ltj mice (Jackson Laboratory, Bar Harbor, Me.) to ensure that theyare normoglycemic (blood glucose is 80-120 mg/dL) prior to initiation ofthe experiment. Blood glucose is measured from tail bleeds using aOneTouch® Ultra® meter and test strips (LifeScan, Milpitas, Calif.).

Cyclophosphamide Induction of Type I Diabetes:

On day 0 and day 14, normoglycemic NOD mice are injectedintraperitoneally with 4 mg cyclophosphamide monohydrate (200 mg/kg)dissolved in 0.9% saline. If mice are diabetic (blood glucose is >250mg/dL), they are not given a booster dose of cyclophosphamide on day 14.

Drug Treatment:

Mice are dosed orally, with vehicle or test compound, once a day fromday 0 until day 25. Compounds are suspended in 0.5% methyl cellulosevehicle using a sonicator to ensure uniform suspension. Mice are weighedtwice weekly and are dosed according to weight. Dosing volume is 5mL/kg. The test compound is dosed at, e.g., 1 mg/kg, 3 mg/kg, 10 mg/kgor 30 mg/kg. Blood glucose is measured twice weekly. After dosing iscompleted at day 25, the mice continue to be monitored and blood glucosemeasurements are taken once a week for 3 weeks. Promotion ofnormoglycemia by the test compound in comparison with vehicle isindicative of the test compound exhibiting therapeutic efficacy in typeI diabetes.

Example 7 Allograft Survival

A compound of the invention can be shown to have therapeutic efficacy inprolonging allograft survival by showing it to have therapeutic efficacyin prolonging, e.g., survival of a skin allograft in an animal model.

Animals:

Female Balbc/J mice (6 to 7 weeks of age at start of study) (JacksonLaboratory, Bar Harbor, Me.) are housed four per cage and maintained ina humidity-controlled (40-60%) and temperature-controlled (68-72° F.)facility on a 12 h:12 h light/dark cycle (lights on at 6:30 am) withfree access to food (Harlan Teklad, Orange, Calif., Rodent Diet 8604)and water. Female C57BL/6 mice (8 to 10 weeks of age at start of study)(Jackson Laboratory, Bar Harbor, Me.) are similarly housed andmaintained. Mice are allowed one week of habituation to the animalfacility before testing.

Skin Allograft:

Balbc/J and C57BL/6 mice are used as donors and recipients,respectively, in a model of skin allograft transplantation. DonorBalbc/J mice are anesthetized, and 0.5 cm-diameter full thickness areasof abdominal skin are surgically removed. Skin grafts harvested from theBalbc/J mice are sutured onto the dorsum of anesthetized recipientC57BL/6 mice. Sutured allografts are covered with Vaseline gauze andBolster dressing for 7 days. The allografted mice are divided into 8groups of 8 mice each.

Clinical Scoring:

Skin allografts are inspected and digital images recorded daily untilrejection, which is defined as the first day on which more than 80% ofthe graft is necrotic. Histological analysis of the rejected graft iscarried out on hematoxylin and eosin (H&E)-stained sections. In anoptional related study, on post-transplantation day 5 isolatedlymphocytes from peripheral lymph nodes and spleen are counted andcharacterized for activation markers (e.g., T-cell activation markers)by flow cytometry. Also on day 5, grafts are removed from transplantedrecipients, cut into small fragments, digested with collagenase andsedimented over Ficoll-Paque (Pharmacia Biotech, Uppsala, Sweden) toisolate graft-infiltrating lymphocytes, which are counted andcharacterized for activation markers (e.g., T-cell activation markers)by flow cytometry. Histological analysis of the graft on day 5 can becarried out on hematoxylin and eosin (H&E)-stained sections.

Drug Treatment:

Mice are dosed orally, with vehicle or test compound, once a day fromthe day of transplantation until the end of the study, e.g. until day14, 21 or 28. Dosing volume is 5 mL/kg. The test compound is dosed at,e.g., 1 mg/kg, 3 mg/kg, 10 mg/kg or 30 mg/kg. Delay of time of rejectionof the skin allograft by the test compound in comparison with vehicle isindicative of the test compound exhibiting therapeutic efficacy inprolonging skin allograft survival.

Example 8 Effect of Compounds on Colitis

A compound of the invention can be shown to have therapeutic efficacy incolitis using an animal model for colitis. Suitable animal models areknown in the art (Boismenu et al., J. Leukoc. Biol., 67:267-278, 2000).A first exemplary animal model for colitis is trinitrobenzenesulfonicacid (TNBS)-induced colitis, which presents clinical andhistopathological findings that resemble those in Crohn's disease(Neurath et al., J. Exp. Med., 182:1281-1290, 1995; Boismenu et al., J.Leukoc. Biol., 67:267-278, 2000). A second exemplary animal model forcolitis is dextran sulfate sodium (DSS)-induced colitis, which presentsclinical and histopathological findings that resemble those inulcerative colitis (Okayasu et al., Gastroenterology, 98:694-702, 1990;Boismenu et al., J. Leukoc. Biol., 67:267-278, 2000). Compounds can becommercially tested for efficacy in at least DSS-induced colitis andTNBS-induced colitis, e.g. by the Jackson Laboratory (Bar Harbor, Me.).

A. Mouse Model for Colitis.

Animals:

Male BALB/c mice (6 weeks of age at start of study) (Jackson Laboratory,Bar Harbor, Me.) are housed four per cage and maintained in ahumidity-controlled (40-60%) and temperature-controlled (68-72° F.)facility on a 12 h:12 h light/dark cycle (lights on at 6:30 am) withfree access to food (Harlan Teklad, Orange Calif., Rodent Diet 8604) andwater. Mice are allowed one week of habituation to the animal facilitybefore testing.

TNBS Induction of Colitis:

Mice are weighed for baseline body weights and fasted later that daybeginning at 6:15 pm just prior to lights-out (day 0). Body weights aretaken again the following morning (day 1) at approximately 7:30 am. Miceare anesthetized with isoflurane prior to induction of colitis. Colitisis induced in the mice by intracolonic injection of about 150 mg/kg TNBSin 50% ethanol (in a volume of 150 μL) using an intubation needle (22 g,1.5 in) inserted completely into the anus with the mouse held by thetail in a vertical position. The mouse is held vertically for 30additional seconds to allow thorough absorption and minimize leakage,after which the mouse is returned to its cage. Mice are then fed,following the preceding approximately 14 hour of fasting. Each morningthereafter, the mice are weighed. In control experiments, mice receive50% ethanol alone using the same protocol.

Drug Treatment:

Drug treatment begins on day 2. Mice are dosed orally, with vehicle or atest compound, once a day from day 2 until the conclusion of theexperiment on, e.g., day 7, 14 or 21. Dosing volume is 5 mL/kg. The testcompound is dosed at, e.g., 1 mg/kg, 3 mg/kg, 10 mg/kg or 30 mg/kg.

Clinical Scoring:

Upon conclusion of the experiment, colons are extracted and measured.Mice are euthanized with CO₂ and colon is removed from anus to cecum.Excised colon is measured for entire length, length from anus to end ofinflamed area and length of inflamed (affected) area. Aftermeasurements, colon is cleared of excrement by flushing with saline andthen cut open to clear more thoroughly. Colon is then weighed andpreserved in neutral buffered formalin (NBF; 10% formalin, pH 6.7-7.0).The colon tissue is embedded in paraffin and processed for hematoxylinand eosin (H & E)-stained sections. Severity of disease symptoms isscored histologically from the stained sections as follows: 0=noevidence of inflammation; 1=low level of leukocyte infiltration withinfiltration seen in <10% of high-power fields AND no structuralchanges; 2=moderate leukocyte infiltration with infiltration seen in 10%to 25% of high-power fields AND crypt elongation AND bowel wallthickening that does not extend beyond the mucosal layer AND noulcerations; 3=high level of leukocyte infiltration seen in 25% to 50%of high-power fields AND crypt elongation AND infiltration beyond themucosal layer AND thickening of the bowel wall AND superficialulcerations; 4=marked degree of transmural leukocyte infiltration seenin >50% of high-power fields AND elongated and distorted crypts ANDbowel wall thickening AND extensive ulcerations. Reduction of theseverity of the disease symptoms by the test compound in comparison withvehicle is indicative of the test compound exhibiting therapeuticefficacy in colitis.

B. Rat Model for Colitis.

Animals:

Male Wistar rats (175-200 g at start of study) (Charles RiverLaboratories, Wilmington, Mass.) are housed two per cage and maintainedin a humidity-controlled (40-60%) and temperature-controlled (68-72° F.)facility on a 12 h:12 h light/dark cycle (lights on at 6:30 am) withfree access to food (Harlan Teklad, Orange Calif., Rodent Diet 8604) andwater. Rats are allowed one week of habituation to the animal facilitybefore testing.

TNBS Induction of Colitis:

Rats are weighed for baseline body weights and fasted later that daybeginning at 6:15 pm just prior to lights-out (day 0). Body weights aretaken again the following morning (day 1) at approximately 7:30 am. Ratsare anesthetized with isoflurane prior to induction of colitis. Colitisis induced in the rats by intracolonic injection of about 60 mg/kg TNBSin 50% ethanol (in a volume of 500 μL) using a fabricated intubationneedle (7.5 Fr umbilical catheter and 14 g hub) inserted 8 cm into theanus with the rat held by the tail in a vertical position. The rat isheld vertically for 30 additional s to allow thorough absorption andminimize leakage, after which the rat is returned to its cage. Rats arethen fed, following the preceding approximately 14 h of fasting. Eachmorning thereafter, the rats are weighed. In control experiments, ratsreceive 50% ethanol alone using the same protocol.

Drug Treatment:

Drug treatment begins on day 2. Rats are dosed orally, with vehicle ortest compound, once a day from day 2 until the conclusion of theexperiment on, e.g., day 7, 14 or 21. Dosing volume is 5 mL/kg. Testcompound is dosed at, e.g., 1 mg/kg, 3 mg/kg, 10 mg/kg or 30 mg/kg.

Clinical Scoring:

Upon conclusion of the experiment, colons are extracted and measured.Rats are euthanized with CO₂ and colon is removed from anus to cecum.Excised colon is measured for entire length, length from anus to end ofinflamed area, and length of inflamed (affected) area. Aftermeasurements, colon is cleared of excrement by flushing with saline andthen cut open to clear more thoroughly. Colon is then weighed andpreserved in neutral buffered formalin (NBF; 10% formalin, pH 6.7-7.0).The colon tissue is embedded in paraffin and processed for hematoxylinand eosin (H & E)-stained sections. Severity of disease symptoms isscored histologically from the stained sections as follows: 0=noevidence of inflammation; 1=low level of leukocyte infiltration withinfiltration seen in <10% of high-power fields AND no structuralchanges; 2=moderate leukocyte infiltration with infiltration seen in 10%to 25% of high-power fields AND crypt elongation AND bowel wallthickening that does not extend beyond the mucosal layer AND noulcerations; 3=high level of leukocyte infiltration seen in 25% to 50%of high-power fields AND crypt elongation AND infiltration beyond themucosal layer AND thickening of the bowel wall AND superficialulcerations; 4=marked degree of transmural leukocyte infiltration seenin >50% of high-power fields AND elongated and distorted crypts ANDbowel wall thickening AND extensive ulcerations. Reduction of theseverity of the disease symptoms by the test compound in comparison withvehicle is indicative of the test compound exhibiting therapeuticefficacy in colitis.

Example 9 Effects of Compounds on Cardiac Telemetry in the Rat

Animals:

Male Sprague-Dawley rats (250-300 g at time of surgery) were implantedby Charles River Laboratories (Wilmington, Mass.) with cardiactransmitting devices (Data Sciences PhysioTel C50-PXT) into theperitoneal space, with a pressure-sensing catheter inserted into thedescending aorta. Rats are allowed at least one week to recover. Ratswere housed in individual cages and maintained in a humidity-controlled(30-70%) and temperature-controlled (20-22° C.) facility on a 12 h:12 hlight/dark cycle (lights on at 7:00 am) with free access to food(Harlan-Teklad, Orange, Calif., Rodent Diet 8604) and water. Rats wereallowed one week of habituation to the animal facility before testing.

Measurement of Cardiovascular Parameters:

The implanted transmitting devices transmitted continuous measurementsof blood pressure (systolic, diastolic, mean arterial, pulse), heartrate, body temperature, and motor activity in freely moving consciousanimals. These data were transmitted via radiofrequency to a computerwhich binned the data into 1 min averages using DataSciences ARTsoftware. Telemetry recording occurred over a 21-h period, starting atnoon and continuing until 9:00 am the following day. A maximum of eightrats were tested at a time, and the same eight rats were utilized forall treatment groups in a within-subject design.

Drug Treatment:

Rats were injected orally with vehicle (PEG400) and the 2^(nd)enantiomer of compound 12 (isolated after resolution of compound 12 byHPLC, with a retention time of 13.9 min per the conditions reported inExample 1.29) at 1:00 pm. A full study (vehicle+3 doses) required fourseparate testing sessions, which occur on Mondays-Tuesdays andThursdays-Fridays. During each of the testing sessions, the eight ratswere divided into four treatment groups such that each group comprisedN=2 for any given session. Rats were re-tested in subsequent testingsessions in a crossover design such that by the end of the foursessions, all animals had received all treatments in a pseudo-randomorder, and each group comprised N=8.

Exemplary Bradycardia Assay:

It was expressly contemplated that the rats could be used to show that acompound of the invention had no or substantially no activity forbradycardia. By way of illustration and not limitation, the rats wereadministered vehicle (PEG 400) and the 2^(nd) enantiomer of compound 12(isolated after resolution of compound 12 by HPLC, with a retention timeof 13.9 min per the conditions reported in Example 1.29) and heart ratewas then measured over a 120 min period. Results are presented in FIG.11. It is apparent from inspection of FIG. 11 that no or substantiallyno reduction of heart rate was exhibited in response to the treatment ofrats with the 2^(nd) enantiomer of compound 12 (isolated afterresolution of compound 12 by HPLC, with a retention time of 13.9 min perthe conditions reported in Example 1.29) in comparison with vehicle. Noor substantially no reduction of heart rate was indicative of the 2^(nd)enantiomer of compound 12 (isolated after resolution of compound 12 byHPLC, with a retention time of 13.9 min per the conditions reported inExample 1.29) exhibiting no or substantially no activity forbradycardia.

Example 10 Effect of Compounds on Arthritis

Female Lewis rats were used in this study. Acclimated animals wereanesthetized with isoflurane and given the first collagen injection (day0). On day 6, they were anesthetized again for the second collageninjection. Collagen was prepared by making a 4 mg/mL solution in 0.01 Nacetic acid. Equal volumes of collagen and incomplete Freund's adjuvantwere emulsified by hand mixing until a bead of this material held itsform when placed in water. Each animal received 300 μL of the mixtureeach time, spread over 3 subcutaneous sites on the back.

Treatment (p.o., q.d., 5 mL/kg dosing volume) began on day 0 andcontinued through day 16 with vehicle or compounds given at 24 hintervals. Rats were weighed on days 0, 3, 6 and 9 through 17 andcaliper measurements of the ankles taken on days 9 through 17. The2^(nd) enantiomer of compound 12 (isolated after resolution of compound12 by HPLC, with a retention time of 13.9 min per the conditionsreported in Example 1.29) was dosed at 0.3, 1 and 3 mg/kg. Results arepresented in FIG. 9. It is apparent from inspection of FIG. 9 that the2^(nd) enantiomer of compound 12 (isolated after resolution of compound12 by HPLC, with a retention time of 13.9 min per the conditionsreported in Example 1.29) exhibited activity for reducing mean anklediameter in the rat.

Example 11 Powder X-Ray Diffraction (PXRD)

Powder X-ray Diffraction (PXRD) data were collected on an XTert PRO MPDpowder diffractometer (PANalytical, Inc.) with a Cu source set at 45 kVand 40 mA, a Ni-filter to remove Cu Kβ radiation, and an X'Celeratordetector. The instrument was calibrated by the vendor using a siliconpowder standard NIST #640c. The calibration was found to be correct whenit was tested with NIST #675 low-angle diffraction standard. Sampleswere prepared for PXRD scanning by placing several milligrams of gentlyground compound onto a sample holder and smoothing as flat as possibleby pressing weigh paper down on the sample with a flat object. Thesamples were analyzed using a spinning-sample stage. Scans cover therange of 5 to 40° 2θ. A continuous scan mode is used with a step size of0.0167° 2θ. Diffraction data were viewed and analyzed with the X'PertData Viewer Software, version 1.0a and X'Pert HighScore Software,version 1.0b

Example 12 Differential Scanning Calorimetry (DSC)

Differential Scanning calorimetry (DSC) was performed on a TAinstruments, Inc. DSC Q2000 at 10° C./min. from ˜25 to ˜210° C. Theinstrument was calibrated at this scan rate by the vendor fortemperature and energy using the melting point and enthalpy of fusion ofan indium standard. Samples were prepared by piercing a sample-pan lidwith a thumb tack or other sharp tool and taring this lid along with asample-pan bottom on a Mettler Toldeo MX5 balance. The sample was placedin the bottom of the tared sample pan. The sample-pan lid fitted snugglyin the sample-pan bottom. The sample and pan were reweighed to get thesample weight. Thermal events (onset temperature, enthalpy of fusion,etc.) were calculated using the Universal Analysis 2000 software,version 4.1D, Build 4.1.0.16.

Example 13 Thermal Gravimetric Analysis (TGA)

Thermal Gravimetric Analysis (TGA) was performed on the TA Instruments,Inc. TGA Q500. The instrument was calibrated by the vendor at 10°C./min. for temperature using the curie point of a ferromagneticstandard. The balance was calibrated with a standard weight. Samplescans were performed at 10° C./min. from 25 to 250° C. Sample was placedinto an open sample pan, previously tared on the TGA balance. Thermalevents such as weight-loss were calculated using the Universal Analysis2000 software, version 4.1D, Build 4.1.0.16.

Example 14 Vapor Sorption Analysis

Hygroscopicity was measured using a dynamic moisture-sorption analyzer,VTI Corporation, SGA-100. The sample was placed as-is in a tared sampleholder on the VTI balance. A drying step was run at 40° C. and 1% RH for20 minutes. The isotherm conditions were 25° C. with steps of 20% RHfrom 10% RH up to 90% RH and back to 10% RH. Weight was checked every 5minutes. Consecutive % weight change of <0.01% or 2 hours, whicheveroccurred first, was required before continuing to the next step.

Those skilled in the art will recognize that various modifications,additions, substitutions and variations to the illustrative examples setforth herein can be made without departing from the spirit of theinvention and are, therefore, considered within the scope of theinvention. All documents referenced above, including, but not limitedto, printed publications and provisional and regular patentapplications, are incorporated herein by reference in their entirety.

1. A process for preparing a composition comprising admixing a compoundselected from compounds of Formula (Ia) and pharmaceutically acceptablesalts, solvates and hydrates thereof:

and a pharmaceutically acceptable carrier; wherein: m is 1; n is 1 or 2;Y is N or CR¹; Z is N or CR⁴; R¹, R², and R⁴ are each independentlyselected from the group consisting of H, C₁-C₆ alkoxy, C₁-C₆ alkyl,C₁-C₆ alkylamino, C₁-C₆ alkylsulfonyl, C₁-C₆ alkylthio, carboxamide,cyano, C₃-C₇ cycloalkoxy, C₃-C₇ cycloalkyl, C₁-C₆ haloalkoxy, C₁-C₆haloalkyl, halogen, heteroaryl and heterocyclyl, wherein said C₁-C₆alkyl and C₁-C₆ alkoxy are each optionally substituted with one or twosubstituents selected from C₃-C₇ cycloalkyl and halogen; and R³ is C₃-C₇cycloalkyl. 2-53. (canceled)
 54. The process according to claim 1,wherein n is
 1. 55. The process according to claim 1, wherein Y is CR¹.56. The process according to claim 55, wherein R¹ is H or C₁-C₆haloalkyl.
 57. The process according to claim 1, wherein R² is selectedfrom the group consisting of H, cyano, C₁-C₆ haloalkoxy and C₁-C₆haloalkyl.
 58. The process according to claim 1, wherein Z is N.
 59. Theprocess according to claim 1, wherein Z is CR⁴.
 60. The processaccording to claim 59, wherein R⁴ is selected from the group consistingof H, cyano, C₁-C₆ haloalkoxy and C₁-C₆ haloalkyl.
 61. The processaccording to claim 1, wherein the compound is selected from compounds ofFormula (Ik) and pharmaceutically acceptable salts, solvates andhydrates thereof:

wherein: Y is N or CR¹; Z is N or CR⁴; R¹ is H or C₁-C₆ haloalkyl; R² isselected from the group consisting of H, cyano, C₁-C₆ haloalkoxy andC₁-C₆ haloalkyl; R³ is C₃-C₇ cycloalkyl; and R⁴ is selected from thegroup consisting of H, cyano, C₁-C₆ haloalkoxy and C₁-C₆ haloalkyl. 62.The process according to claim 1, wherein the compound is selected fromcompounds of Formula (Ik) and pharmaceutically acceptable salts,solvates and hydrates thereof:

wherein: Y is N or CR¹; Z is N or CR⁴; R¹ is H or trifluoromethyl; R² isselected from the group consisting of H, cyano, trifluoromethoxy andtrifluoromethyl; R³ is selected from the group consisting of cyclobutyl,cyclohexyl, cyclopentyl, and cyclopropyl; and R⁴ is selected from thegroup consisting of H, cyano, trifluoromethoxy and trifluoromethyl. 63.The process according to claim 1, wherein the compound is selected fromcompounds of Formula (Im) and pharmaceutically acceptable salts,solvates and hydrates thereof:

wherein: R² is selected from the group consisting of H, cyano,trifluoromethoxy and trifluoromethyl; and R³ is selected from the groupconsisting of cyclobutyl, cyclohexyl, cyclopentyl, and cyclopropyl. 64.The process according to claim 1, wherein the compound is selected fromthe following compounds and pharmaceutically acceptable salts, solvatesand hydrates thereof:2-(7-(4-cyclohexyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)aceticacid;(R)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)aceticacid;(S)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)aceticacid;2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)aceticacid;2-(7-(3-cyano-4-cyclohexylbenzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3yl)aceticacid;2-(7-((6-cyclopentyl-5-(trifluoromethyl)pyridin-3-yl)methoxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)aceticacid;2-(7-(4-cyclobutyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)aceticacid; and2-(7-(4-cyclopropyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)aceticacid.
 65. The process according to claim 1, wherein the compound isselected from the following compound and pharmaceutically acceptablesalts, solvates and hydrates thereof:(R)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)aceticacid.
 66. The process according to claim 1, wherein the compound isselected from the following compound and pharmaceutically acceptablesalts, solvates and hydrates thereof:(S)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)aceticacid.
 67. A process according to claim 1, wherein the salt is selectedfrom the following salt and pharmaceutically acceptable solvates andhydrates thereof: L-Arginine salt of(R)-2-(7-(4-cyclopentyl-3-(trifluoromethyl)benzyloxy)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yl)aceticacid.
 68. The process according to claim 1, wherein the composition issuitable for oral, rectal, nasal, topical, buccal, sub-lingual, vaginal,parenteral, intramuscular, subcutaneous, or intravenous administration;or suitable for administration by inhalation, insufflation or by atransdermal patch.
 69. The process according to claim 1, wherein thecomposition is suitable for oral administration.
 70. The processaccording to claim 1, wherein the pharmaceutically acceptable carriercomprises a diluent.
 71. The process according to claim 1, wherein thepharmaceutically acceptable carrier comprises mannitol.
 72. The processaccording to claim 1, wherein the pharmaceutically acceptable carriercomprises a lubricant.
 73. The process according to claim 1, wherein thepharmaceutically acceptable carrier comprises magnesium stearate. 74.The process according to claim 1, wherein the pharmaceuticallyacceptable carrier comprises a tablet disintegrating agent.
 75. Theprocess according to claim 1, wherein the pharmaceutically acceptablecarrier comprises a diluent, a lubricant, and a tablet disintegratingagent.
 76. The process according to claim 1, wherein thepharmaceutically acceptable carrier comprises mannitol and magnesiumstearate.
 77. The process according to claim 65, wherein the compositionis suitable for oral, rectal, nasal, topical, buccal, sub-lingual,vaginal, parenteral, intramuscular, subcutaneous, or intravenousadministration; or suitable for administration by inhalation,insufflation or by a transdermal patch.
 78. The process according toclaim 65, wherein the composition is suitable for oral administration.79. The process according to claim 65, wherein the pharmaceuticallyacceptable carrier comprises a diluent.
 80. The process according toclaim 65, wherein the pharmaceutically acceptable carrier comprisesmannitol.
 81. The process according to claim 65, wherein thepharmaceutically acceptable carrier comprises a lubricant.
 82. Theprocess according to claim 65, wherein the pharmaceutically acceptablecarrier comprises magnesium stearate.
 83. The process according to claim65, wherein the pharmaceutically acceptable carrier comprises a tabletdisintegrating agent.
 84. The process according to claim 65, wherein thepharmaceutically acceptable carrier comprises a diluent, a lubricant,and a tablet disintegrating agent.
 85. The process according to claim65, wherein the pharmaceutically acceptable carrier comprises mannitoland magnesium stearate.
 86. The process according to claim 67, whereinthe composition is suitable for oral, rectal, nasal, topical, buccal,sub-lingual, vaginal, parenteral, intramuscular, subcutaneous, orintravenous administration; or suitable for administration byinhalation, insufflation or by a transdermal patch.
 87. The processaccording to claim 67, wherein the composition is suitable for oraladministration.
 88. The process according to claim 67, wherein thepharmaceutically acceptable carrier comprises a diluent.
 89. The processaccording to claim 67, wherein the pharmaceutically acceptable carriercomprises mannitol.
 90. The process according to claim 67, wherein thepharmaceutically acceptable carrier comprises a lubricant.
 91. Theprocess according to claim 67, wherein the pharmaceutically acceptablecarrier comprises magnesium stearate.
 92. The process according to claim67, wherein the pharmaceutically acceptable carrier comprises a tabletdisintegrating agent.
 93. The process according to claim 67, wherein thepharmaceutically acceptable carrier comprises a diluent, a lubricant,and a tablet disintegrating agent.
 94. The process according to claim67, wherein the pharmaceutically acceptable carrier comprises mannitoland magnesium stearate.